LOCUS       Exported                6269 bp DNA     circular SYN 08-FEB-2019
DEFINITION  pET-21 b (+) with PETcan_101 sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6269)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 27, 2021 from SnapGene 5.3.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6269
                     /organism="pET-21 b (+)"
                     /mol_type="other DNA"
                     /label=source:pET-21 b (+)
     variation       458
                     /label=T->C
     variation       467..468
                     /label=TCA->TA
     promoter        528..556
                     /label=AmpR_promoter
                     /note="/translation=MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLG
                     ARVGY 
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE 
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL 
                     DRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL 
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA 
                     EIGASLIKHW*"
     CDS             598..1458
                     /codon_start=1
                     /label=ORF frame 2(1)
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            598..1458
                     /gene="Ampicillin"
                     /label=Ampicillin
                     /note="/translation=MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIV
                     AAIKS 
                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV 
                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW 
                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE 
                     SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ 
                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
     rep_origin      1613..2232
                     /label=pBR322_origin
                     /note="/translation=MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIV
                     AAIKS 
                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV 
                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW 
                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE 
                     SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ 
                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
     variation       2452..2453
                     /label=ATATA->AA
     misc_feature    2628..2650
                     /label=pGEX_3_primer
                     /note="/translation=MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIV
                     AAIKS 
                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV 
                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW 
                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE 
                     SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ 
                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
     misc_feature    complement(2644..2835)
                     /label=ROP
                     /note="/translation=MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIV
                     AAIKS 
                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV 
                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW 
                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE 
                     SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ 
                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
     misc_feature    complement(3644..4735)
                     /label=lacI
                     /note="/translation=MSACFGVGMVAGPVAGGLLGAISLHAPFLAAAVLNGLNL
                     LLGCF 
                     LMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGECETSNVIRCRRVCRCLLSD 
                     RFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQTVVADWRCHLQSGPARAV 
                     ANCRGD*"
     CDS             complement(3644..4603)
                     /codon_start=1
                     /label=ORF frame 2
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSR
                     ADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPA
                     LFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKY
                     LTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLR
                     VGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVS
                     LVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ"
     CDS             complement(4476..4976)
                     /codon_start=1
                     /label=ORF frame 1
                     /translation="MSACFGVGMVAGPVAGGLLGAISLHAPFLAAAVLNGLNLLLGCFL
                     MQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGECETSNVIRCRRVCRCLLSDRF
                     PRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQTVVADWRCHLQSGPARAVANC
                     RGD"
     misc_feature    5010..5029
                     /label=pBRrevBam_primer
     primer_bind     5113..5132
                     /label=T7 primer
     promoter        5113..5131
                     /label=T7_promoter
     misc_feature    5131..5158
                     /label=lacO
     misc_feature    5176..5192
                     /label=T7_transl_en_RBS
     CDS             5201..6112
                     /codon_start=1
                     /label=PETcan_101
                     /translation="MCLYLNIWTPDLNGSLPVMVFIHGGGNQQGSTAQIAGGARIYEGK
                     NLARRGQVVVVTLQYRLGALGYLVHPGLEAESTHGKAGNYGALDQLAALLWIKENIRAF
                     GGDPELVTLFGESAGAVNIGNLLVMPAAKGLFHRAILQSGSPRLKAYSAARNEGIAFAQ
                     KLGAAGTPEQQVAHLRTLPVDSLVKGDSNPISGGSMAQGSWQPVLDGYWFPQAPLDAMR
                     SGEHHRVPLIVGSSSDEMSLYVPSVVTPLMLQTFVQTTIPAPYRQQVLALYPPGTTNEQ
                     ARASYVALVGDPLESTCRHASLE"
     misc_signal     6113..6130
                     /label=His Tag
     terminator      6141..6269
                     /label=T7_terminator
     primer_bind     complement(6183..6201)
                     /label=T7 Term primer
ORIGIN
        1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
       61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
      121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
      181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
      241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
      301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
      361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
      421 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt
      481 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
      541 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
      601 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
      661 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
      721 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
      781 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
      841 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
      901 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
      961 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     1021 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     1081 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     1141 gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     1201 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     1261 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     1321 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     1381 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     1441 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     1501 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     1561 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     1621 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     1681 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     1741 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     1801 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     1861 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     1921 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     1981 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     2041 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     2101 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     2161 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     2221 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     2281 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     2341 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     2401 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg caatggtgca
     2461 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct
     2521 acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg
     2581 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     2641 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc
     2701 agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag
     2761 tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt
     2821 ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat
     2881 accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt
     2941 actggaacgt tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat
     3001 cactcagggt caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca
     3061 gcagcatcct gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc
     3121 cagactttac gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt
     3181 tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt
     3241 aaggcaaccc cgccagccta gccgggtcct caacgacagg agcacgatca tgcgcacccg
     3301 tggggccgcc atgccggcga taatggcctg cttctcgccg aaacgtttgg tggcgggacc
     3361 agtgacgaag gcttgagcga gggcgtgcaa gattccgaat accgcaagcg acaggccgat
     3421 catcgtcgcg ctccagcgaa agcggtcctc gccgaaaatg acccagagcg ctgccggcac
     3481 ctgtcctacg agttgcatga taaagaagac agtcataagt gcggcgacga tagtcatgcc
     3541 ccgcgcccac cggaaggagc tgactgggtt gaaggctctc aagggcatcg gtcgagatcc
     3601 cggtgcctaa tgagtgagct aacttacatt aattgcgttg cgctcactgc ccgctttcca
     3661 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     3721 tttgcgtatt gggcgccagg gtggtttttc ttttcaccag tgagacgggc aacagctgat
     3781 tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca
     3841 gcaggcgaaa atcctgtttg atggtggtta acggcgggat ataacatgag ctgtcttcgg
     3901 tatcgtcgta tcccactacc gagatatccg caccaacgcg cagcccggac tcggtaatgg
     3961 cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagtg ggaacgatgc
     4021 cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc
     4081 gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca
     4141 gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg
     4201 cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga
     4261 tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca
     4321 cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg acgcgttgcg
     4381 cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca
     4441 ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg
     4501 gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca
     4561 gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt
     4621 cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag
     4681 agacaccggc atactctgcg acatcgtata acgttactgg tttcacattc accaccctga
     4741 attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg
     4801 tgtccgggat ctcgacgctc tcccttatgc gactcctgca ttaggaagca gcccagtagt
     4861 aggttgaggc cgttgagcac cgccgccgca aggaatggtg catgcaagga gatggcgccc
     4921 aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc
     4981 ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc
     5041 gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggat cgagatctcg
     5101 atcccgcgaa attaatacga ctcactatag gggaattgtg agcggataac aattcccctc
     5161 tagaaataat tttgtttaac tttaagaagg agatatacat atgtgcctgt atctgaatat
     5221 ctggactccg gacctgaatg gcagcctgcc ggttatggtg ttcattcacg gcggcggtaa
     5281 ccagcaaggt tccaccgctc agatcgccgg tggtgcacgt atctacgagg gcaagaacct
     5341 ggcacgtcgt ggtcaggtag ttgtagtcac ccttcaatac cgtctgggtg cactgggcta
     5401 tctggtacac ccgggtctgg aagctgaatc cacccacggc aaggctggta actatggggc
     5461 tctggatcag ctggcagcac tgctctggat taaagaaaac atccgcgctt ttggtggtga
     5521 cccggagctg gtcacgctgt tcggcgaaag cgcgggcgcg gttaacatcg gcaacctgct
     5581 ggttatgccg gctgcaaaag gtctgtttca ccgtgctatc ctgcagtcgg ggtccccgcg
     5641 cctgaaagcc tattctgcag cgcgcaacga aggtatcgct tttgcacaga agctgggtgc
     5701 tgcaggtact ccagagcagc aggtggccca tctgcgcact ctgcctgtcg atagcctggt
     5761 gaaaggcgat tctaacccga tcagcggcgg cagcatggcg cagggtagct ggcagccggt
     5821 actggatggt tactggttcc cacaggctcc gctcgatgcg atgcgtagcg gtgaacacca
     5881 tcgtgtaccg ctgatcgttg gttcttcctc ggacgagatg agcctgtacg ttccgtctgt
     5941 tgtgactccg ctgatgctgc agacttttgt tcagaccact attccggcgc cttatcgcca
     6001 gcaggtcctg gcactgtacc cgccgggtac caccaacgaa caagcccgtg catcttacgt
     6061 ggccctggtc ggtgacccgc tggagtccac ttgtcgtcac gcaagcctcg agcaccacca
     6121 ccaccaccac tgagatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc
     6181 caccgctgag caataactag cataacccct tggggcctct aaacgggtct tgaggggttt
     6241 tttgctgaaa ggaggaacta tatccggat
//
