LOCUS       Exported                7532 bp ds-DNA     circular SYN 16-MAR-2019
DEFINITION  Bacterial vector for the co-expression of two genes.
ACCESSION   .
VERSION     .
KEYWORDS    pETDuet-1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7532)
  AUTHORS   MilliporeSigma (Novagen)
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Apr 20, 2021 from SnapGene Viewer 5.2.4
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7532
                     /organism="synthetic DNA construct"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
     protein_bind    3..27
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /note="color: #31849b"
     RBS             58..63
                     /note="ribosome binding site"
                     /note="color: #a6acb3"
     CDS             71..73
                     /codon_start=1
                     /product="start codon"
                     /label=ATG
                     /note="color: #993366"
                     /translation="M"
     CDS             83..100
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /note="color: #cc99b2"
                     /translation="HHHHHH"
     misc_feature    112
                     /label=INSERT
                     /note="color: #a6acb3"
     CDS             113..1432
                     /codon_start=1
                     /label=alpha
                     /note="color: #993366"
                     /translation="MADPDVLTEVPAALKRLAKYVIRGFYGIEHALALDILIRNSCVKE
                     EDMLELLKFDRKQLRSVLNNLKGDKFIKCRMRVETAADGKTTRHNYYFINYRTLVNVVK
                     YKLDHMRRRIETDERDSTNRASFKCPVCSSTFTDLEANQLFDPMTGTFRCTFCHTEVEE
                     DESAMPKKDARTLLARFNEQIEPIYALLRETEDVNLAYEILEPEPTEIPALKQSKDHAA
                     TTAGAASLAGGHHREAWATKGPSYEDLYTQNVVINMDDQEDLHRASLEGKSAKERPIWL
                     RESTVQGAYGSEDMKEGGIDMDAFQEREEGHAGPDDNEEVMRALLIHEKKTSSAMAGSV
                     GAAAPVTAANGSDSESETSESDDDSPPRPAAVAVHKREEDEEEDDEFEEVADDPIVMVA
                     GRPFSYSEVSQRPELVAQMTPEEKEAYIAMGQRMFEDLFE"
     primer_bind     1479..1498
                     /label=DuetUP2 Primer
                     /note="color: black; sequence: TTGTACACGGCCGCATAATC"
     primer_bind     complement(1479..1498)
                     /label=DuetDOWN1 Primer
                     /note="color: black; sequence: GATTATGCGGCCGTGTACAA"
     promoter        1504..1522
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     protein_bind    1523..1547
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /note="color: #31849b"
     RBS             1576..1581
                     /note="ribosome binding site"
                     /note="color: #a6acb3"
     CDS             1590..2465
                     /codon_start=1
                     /label=beta
                     /note="color: #993366"
                     /translation="MDPSLLRERELFKKRALSTPVVEKRSASSESSSSSSKKKKTKVEH
                     GGSSGSKQNSDHSNGSFNLKALSGSSGYKFGVLAKIVNYMKTRHQRGDTHPLTLDEILD
                     ETQHLDIGLKQKQWLMTEALVNNPKIEVIDGKYAFKPKYNVRDKKALLRLLDQHDQRGL
                     GGILLEDIEEALPNSQKAVKALGDQILFVNRPDKKKILFFNDKSCQFSVDEEFQKLWRS
                     VTVDSMDEEKIEEYLKRQGISSMQESGPKKVAPIQRRKKPASQKKRRFKTHNEHLAGVL
                     KDYSDITSSK"
     CDS             2478..2522
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /note="color: #cc99b2"
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(2560..2578)
                     /label=T7 Terminator Primer
                     /note="color: black; sequence: GCTAGTTATTGCTCAGCGG"
     terminator      2574..2621
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
                     /note="color: #ffffff"
     rep_origin      2658..3113
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     CDS             complement(3228..4088)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 3228 .. 4019 / #ccffcc
                        2: 4020 .. 4088 / #ccffcc / signal sequence
                     Cleavage site after base 4019"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4089..4181)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
     rep_origin      4262..4850
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(5280..5471)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /note="color: #993366"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(6043..7125)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /note="color: #993366"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(7126..7203)
                     /gene="lacI"
                     /label=lacI promoter
                     /note="color: #ffffff; direction: LEFT"
     primer_bind     7470..7485
                     /label=pET Upstream Primer
                     /note="color: black; sequence: ATGCGTCCGGCGTAGA"
     promoter        join(7516..7532,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag
       61 gagatatacc atgggcagca gccatcacca tcatcaccac agccaggatc ccatggcaga
      121 cccagatgtc ctcactgaag ttccagcagc attgaagcgg ttagccaagt atgtgatccg
      181 gggattttat ggcattgagc atgccttggc cttggacatc ttgatcagga actcctgtgt
      241 gaaagaggag gatatgctgg agctgctcaa gtttgatcgg aagcaacttc gatcagtttt
      301 gaataattta aagggagaca agtttatcaa atgcagaatg agggtagaga ctgctgcaga
      361 cgggaaaacc actcgccata actactactt catcaattat cgtactcttg ttaatgtggt
      421 aaaatataaa ctggaccaca tgagaagaag aattgagacc gatgagagag attcgaccaa
      481 ccgggcttcc ttcaaatgtc ctgtctgtag tagtactttc acagacttag aagctaatca
      541 gctctttgat cctatgacag gaactttccg ctgtactttt tgccatacag aggtagaaga
      601 ggatgaatca gcaatgccca aaaaagatgc acgcacactt ttggcaaggt ttaatgaaca
      661 aattgagccc atttatgcat tgcttcggga gacagaggat gtgaacttgg cctatgaaat
      721 acttgagcca gaacccacag aaatcccagc cctgaaacag agcaaggacc atgcagcaac
      781 tactgctgga gctgctagcc tagcaggtgg gcaccaccgg gaagcatggg ccaccaaagg
      841 tccttcctat gaagacttat acactcagaa tgttgtcatt aacatggatg accaagaaga
      901 tcttcatcga gcctcactgg aagggaaatc tgccaaagag aggcctattt ggttgagaga
      961 aagcactgtc caaggggcat atggttctga agatatgaaa gaagggggca tagatatgga
     1021 cgcatttcag gagcgtgagg aaggccatgc tgggcctgat gacaacgaag aggtcatgcg
     1081 agcactgctc attcacgaga aaaagacttc ctctgccatg gctggttcag tgggggcagc
     1141 tgctccagtg accgctgcca atggcagtga ctcagaaagc gagaccagtg agtcagatga
     1201 tgattctcca ccccgtccgg cagctgtggc tgtgcataaa cgagaagagg atgaagagga
     1261 agatgacgag tttgaagaag tagcagatga ccccattgtc atggtggctg gccgtccgtt
     1321 ctcctacagt gaagtgagcc aacggccaga gctagtggcc cagatgacac cagaagaaaa
     1381 ggaagcatat atagcaatgg gacaacgcat gtttgaggac ctctttgagt gaaagcttgc
     1441 ggccgcataa tgcttaagtc gaacagaaag taatcgtatt gtacacggcc gcataatcga
     1501 aattaatacg actcactata ggggaattgt gagcggataa caattcccca tcttagtata
     1561 ttagttaagt ataagaagga gatatacata tggatccaag cctgttgaga gaaagggagc
     1621 tgttcaaaaa acgagctctt tctactcctg tagtagaaaa acgttcagca tcttctgagt
     1681 catcatcatc atcgtcaaag aagaagaaaa caaaggtaga acatggagga tcgtcaggct
     1741 ctaaacaaaa ttctgatcat agcaatggat catttaactt gaaagctttg tcaggaagct
     1801 ctggatataa gtttggtgtt cttgctaaga ttgtgaatta catgaagaca cggcatcagc
     1861 gaggagatac gcatcctcta accttagatg aaattttgga tgaaacacaa catttagata
     1921 ttggactcaa gcagaaacaa tggctaatga ctgaggcttt agtcaacaat cccaaaattg
     1981 aagtaataga tgggaagtat gctttcaagc ccaagtacaa cgtgagagat aagaaggccc
     2041 tacttaggct cttagatcag catgaccagc gaggattagg aggaattctt ttagaagaca
     2101 tagaagaagc actgcccaat tcccagaaag ctgtcaaggc tttgggggac cagatactat
     2161 ttgtaaatcg tcccgataag aagaaaatac ttttcttcaa tgataagagc tgtcagtttt
     2221 ctgtggatga agaatttcag aaactgtgga ggagtgtcac tgtagattcc atggacgagg
     2281 agaaaattga agaatatctg aagcgacagg gtatttcttc catgcaggaa tctggaccaa
     2341 agaaagtggc ccctattcag agaaggaaaa agcctgcttc acagaaaaag cgacgcttta
     2401 agactcataa cgaacacttg gctggagtgc tgaaggatta ctctgacatt acttccagca
     2461 aatagctcga gtctggtaaa gaaaccgctg ctgcgaaatt tgaacgccag cacatggact
     2521 cgtctactag cgcagcttaa ttaacctagg ctgctgccac cgctgagcaa taactagcat
     2581 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat
     2641 ccggattggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     2701 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     2761 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     2821 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     2881 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     2941 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     3001 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     3061 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa tttctggcgg
     3121 cacgatggca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     3181 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     3241 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     3301 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     3361 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     3421 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     3481 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     3541 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     3601 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     3661 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     3721 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     3781 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     3841 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     3901 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     3961 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     4021 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     4081 atactcatac tcttcctttt tcaatcatga ttgaagcatt tatcagggtt attgtctcat
     4141 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggtcatg accaaaatcc
     4201 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     4261 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     4321 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     4381 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     4441 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     4501 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     4561 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     4621 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     4681 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     4741 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     4801 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     4861 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     4921 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     4981 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga
     5041 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatat ggtgcactct
     5101 cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt
     5161 gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     5221 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     5281 cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg
     5341 tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc
     5401 tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc
     5461 tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt aatgataccg
     5521 atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg
     5581 gaacgttgtg agggtaaaca actggcggta tggatgcggc gggaccagag aaaaatcact
     5641 cagggtcaat gccagcgctt cgttaataca gatgtaggtg ttccacaggg tagccagcag
     5701 catcctgcga tgcagatccg gaacataatg gtgcagggcg ctgacttccg cgtttccaga
     5761 ctttacgaaa cacggaaacc gaagaccatt catgttgttg ctcaggtcgc agacgttttg
     5821 cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta accagtaagg
     5881 caaccccgcc agcctagccg ggtcctcaac gacaggagca cgatcatgct agtcatgccc
     5941 cgcgcccacc ggaaggagct gactgggttg aaggctctca agggcatcgg tcgagatccc
     6001 ggtgcctaat gagtgagcta acttacatta attgcgttgc gctcactgcc cgctttccag
     6061 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     6121 ttgcgtattg ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt
     6181 gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag
     6241 caggcgaaaa tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt
     6301 atcgtcgtat cccactaccg agatgtccgc accaacgcgc agcccggact cggtaatggc
     6361 gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc
     6421 ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg
     6481 ttccgctatc ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag
     6541 acgcgccgag acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc
     6601 gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat
     6661 gggtgtctgg tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac
     6721 agcaatggca tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc
     6781 gagaagattg tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac
     6841 caccacgctg gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg
     6901 cgcgtgcagg gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag
     6961 ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc
     7021 ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga
     7081 gacaccggca tactctgcga catcgtataa cgttactggt ttcacattca ccaccctgaa
     7141 ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcgcc attcgatggt
     7201 gtccgggatc tcgacgctct cccttatgcg actcctgcat taggaagcag cccagtagta
     7261 ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag atggcgccca
     7321 acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg ctcatgagcc
     7381 cgaagtggcg agcccgatct tccccatcgg tgatgtcggc gatataggcg ccagcaaccg
     7441 cacctgtggc gccggtgatg ccggccacga tgcgtccggc gtagaggatc gagatcgatc
     7501 tcgatcccgc gaaattaata cgactcacta ta
//
