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pETDuet1載體基本信息
| EMD Biosciences (Novagen) |
| pETDuet1, pETDuet 1, pETDuet-1 |
| 大腸桿菌蛋白雙表達(dá)載體，有兩個多克隆位點 |
| 高 |
| 多克隆位點，限制性內(nèi)切酶 |
| 5420 bp |
| pET Upstream Primer: 5'-ATGCGTCCGGCGTAGA-3' |
| DuetDOWN1 Primer: 5'-GATTATGCGGCCGTGTACAA-3' |
| DuetUP2 Primer: 5'-TTGTACACGGCCGCATAATC-3' |
| T7t: 5' GCTAGTTATTGCTCAGCGG 3' |
| N-His, C-S |
| 氨芐 |
pETDuet-1是設(shè)計用來同時表達(dá)兩個目的基因。pETDuet-1攜帶有pBR322質(zhì)粒來源的ColE1復(fù)制子, lacI基因和氨芐抗性基因。 |
| 71146-3 |
| 瞬時表達(dá) |
| 組成型 |
| 非病毒 |
pETDuet1載體質(zhì)粒圖譜和多克隆位點信息
pETDuet1載體簡介
pETDuet-1載體被設(shè)計用來在大腸桿菌中同時表達(dá)兩個外源基因. 載體含有兩個多克隆位點，每一個多克隆位點是通過T7啟動子/Lac操縱子和一個核糖體結(jié)合位點組成。pETDuet-1攜帶有pBR322質(zhì)粒來源的ColE1復(fù)制子, lacI基因和氨芐抗性基因。
本載體可以與pACYCDuet-1載體聯(lián)合使用，在合適的宿主菌中，可以一起共表達(dá)至多4個目的基因。插入在pETDuet-1載體多克隆位點1的目的基因可以使用pET Upstream Primer (Cat. No. 69214-3)和DuetDOWN1 Primer (Cat. No. 71179-3)進(jìn)行測序，插入在多克隆位點2的目的基因可以通過DuetUP2 Primer (Cat. No. 71180-3)和T7 Terminator Primer (Cat. No. 69337-3)進(jìn)行目的基因測序。
pETDuet1載體序列
LOCUS pETDuet-1 5420 bp DNA circular 7-SEP-2008
SOURCE
ORGANISM
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT ORIGDB|GenBank
COMMENT VNTAUTHORNAME|biofeng.com|
COMMENT VNTOAUTHORAD1|Novagen Inc.|
COMMENT VNTREPLTYPE|Plasmid
FEATURES Location/Qualifiers
promoter 5404..5420
/vntifkey="30"
/label=T7\promoter-1
misc_feature 1..1
/vntifkey="21"
/label=T7\transcription\start-1
misc_feature 83..100
/vntifkey="21"
/label=His-Tag
misc_feature 70..168
/vntifkey="21"
/label=MCS-1
promoter 214..230
/vntifkey="30"
/label=T7\promoter-2
misc_feature 231..231
/vntifkey="21"
/label=T7\transcription\start-2
misc_feature 297..438
/vntifkey="21"
/label=MCS-2
misc_feature 366..410
/vntifkey="21"
/label=S-Tag
terminator 462..509
/vntifkey="43"
/label=T7\terminator
operon 3931..5013
/vntifkey="103"
/label=lacI\operon
rep_origin 2737..2737
/vntifkey="33"
/label=pBR322\origin
misc_feature 1119..1976
/vntifkey="21"
/label=Amp
rep_origin 547..994
/vntifkey="33"
/label=f1\origin
BASE COUNT 1274 a 1434 c 1400 g 1312 t
ORIGIN
1 ggggaattgt gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag
61 gagatatacc atgggcagca gccatcacca tcatcaccac agccaggatc cgaattcgag
121 ctcggcgcgc ctgcaggtcg acaagcttgc ggccgcataa tgcttaagtc gaacagaaag
181 taatcgtatt gtacacggcc gcataatcga aattaatacg actcactata ggggaattgt
241 gagcggataa caattcccca tcttagtata ttagttaagt ataagaagga gatatacata
301 tggcagatct caattggata tcggccggcc acgcgatcgc tgacgtcggt accctcgagt
361 ctggtaaaga aaccgctgct gcgaaatttg aacgccagca catggactcg tctactagcg
421 cagcttaatt aacctaggct gctgccaccg ctgagcaata actagcataa ccccttgggg
481 cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggattggcga
541 atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
601 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
661 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
721 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
781 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
841 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
901 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
961 atttaacgcg aattttaaca aaatattaac gtttacaatt tctggcggca cgatggcatg
1021 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
1081 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
1141 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
1201 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
1261 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
1321 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
1381 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
1441 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
1501 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
1561 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
1621 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
1681 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
1741 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
1801 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
1861 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
1921 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
1981 ttcctttttc aatcatgatt gaagcattta tcagggttat tgtctcatga gcggatacat
2041 atttgaatgt atttagaaaa ataaacaaat aggtcatgac caaaatccct taacgtgagt
2101 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
2161 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
2221 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
2281 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
2341 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
2401 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
2461 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
2521 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
2581 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
2641 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
2701 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
2761 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
2821 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
2881 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc
2941 tccttacgca tctgtgcggt atttcacacc gcatatatgg tgcactctca gtacaatctg
3001 ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg
3061 gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
3121 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
3181 ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc
3241 gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt
3301 aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact
3361 gatgcctccg tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag
3421 aggatgctca cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag
3481 ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc
3541 cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg
3601 cagatccgga acataatggt gcagggcgct gacttccgcg tttccagact ttacgaaaca
3661 cggaaaccga agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg
3721 cttcacgttc gctcgcgtat cggtgattca ttctgctaac cagtaaggca accccgccag
3781 cctagccggg tcctcaacga caggagcacg atcatgctag tcatgccccg cgcccaccgg
3841 aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg tgcctaatga
3901 gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
3961 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
4021 cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc
4081 ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc
4141 ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc
4201 cactaccgag atgtccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc
4261 cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat
4321 ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg
4381 ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac
4441 agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc
4501 cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc
4561 agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc
4621 ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg
4681 caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc
4741 acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc
4801 cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac
4861 gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc
4921 agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata
4981 ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc
5041 cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc
5101 gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt
5161 tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg
5221 ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag
5281 cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc
5341 cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatcgatctc gatcccgcga
5401 aattaatacg actcactata
//
SOURCE
ORGANISM
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT ORIGDB|GenBank
COMMENT VNTAUTHORNAME|biofeng.com|
COMMENT VNTOAUTHORAD1|Novagen Inc.|
COMMENT VNTREPLTYPE|Plasmid
FEATURES Location/Qualifiers
promoter 5404..5420
/vntifkey="30"
/label=T7\promoter-1
misc_feature 1..1
/vntifkey="21"
/label=T7\transcription\start-1
misc_feature 83..100
/vntifkey="21"
/label=His-Tag
misc_feature 70..168
/vntifkey="21"
/label=MCS-1
promoter 214..230
/vntifkey="30"
/label=T7\promoter-2
misc_feature 231..231
/vntifkey="21"
/label=T7\transcription\start-2
misc_feature 297..438
/vntifkey="21"
/label=MCS-2
misc_feature 366..410
/vntifkey="21"
/label=S-Tag
terminator 462..509
/vntifkey="43"
/label=T7\terminator
operon 3931..5013
/vntifkey="103"
/label=lacI\operon
rep_origin 2737..2737
/vntifkey="33"
/label=pBR322\origin
misc_feature 1119..1976
/vntifkey="21"
/label=Amp
rep_origin 547..994
/vntifkey="33"
/label=f1\origin
BASE COUNT 1274 a 1434 c 1400 g 1312 t
ORIGIN
1 ggggaattgt gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag
61 gagatatacc atgggcagca gccatcacca tcatcaccac agccaggatc cgaattcgag
121 ctcggcgcgc ctgcaggtcg acaagcttgc ggccgcataa tgcttaagtc gaacagaaag
181 taatcgtatt gtacacggcc gcataatcga aattaatacg actcactata ggggaattgt
241 gagcggataa caattcccca tcttagtata ttagttaagt ataagaagga gatatacata
301 tggcagatct caattggata tcggccggcc acgcgatcgc tgacgtcggt accctcgagt
361 ctggtaaaga aaccgctgct gcgaaatttg aacgccagca catggactcg tctactagcg
421 cagcttaatt aacctaggct gctgccaccg ctgagcaata actagcataa ccccttgggg
481 cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggattggcga
541 atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
601 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
661 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
721 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
781 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
841 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
901 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
961 atttaacgcg aattttaaca aaatattaac gtttacaatt tctggcggca cgatggcatg
1021 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
1081 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
1141 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
1201 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
1261 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
1321 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
1381 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
1441 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
1501 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
1561 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
1621 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
1681 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
1741 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
1801 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
1861 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
1921 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
1981 ttcctttttc aatcatgatt gaagcattta tcagggttat tgtctcatga gcggatacat
2041 atttgaatgt atttagaaaa ataaacaaat aggtcatgac caaaatccct taacgtgagt
2101 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
2161 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
2221 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
2281 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
2341 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
2401 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
2461 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
2521 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
2581 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
2641 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
2701 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
2761 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
2821 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
2881 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc
2941 tccttacgca tctgtgcggt atttcacacc gcatatatgg tgcactctca gtacaatctg
3001 ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg
3061 gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
3121 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
3181 ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc
3241 gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt
3301 aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact
3361 gatgcctccg tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag
3421 aggatgctca cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag
3481 ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc
3541 cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg
3601 cagatccgga acataatggt gcagggcgct gacttccgcg tttccagact ttacgaaaca
3661 cggaaaccga agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg
3721 cttcacgttc gctcgcgtat cggtgattca ttctgctaac cagtaaggca accccgccag
3781 cctagccggg tcctcaacga caggagcacg atcatgctag tcatgccccg cgcccaccgg
3841 aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg tgcctaatga
3901 gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
3961 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
4021 cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc
4081 ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc
4141 ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc
4201 cactaccgag atgtccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc
4261 cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat
4321 ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg
4381 ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac
4441 agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc
4501 cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc
4561 agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc
4621 ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg
4681 caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc
4741 acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc
4801 cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac
4861 gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc
4921 agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata
4981 ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc
5041 cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc
5101 gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt
5161 tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg
5221 ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag
5281 cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc
5341 cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatcgatctc gatcccgcga
5401 aattaatacg actcactata
//
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