LOCUS       sequence-443290-        6067 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6067)
  AUTHORS   Yasugi T, Benson JD, Sakai H, Vidal M, Howley PM
  TITLE     Mapping and characterization of the interaction domains of human 
            papillomavirus type 16 E1 and E2 proteins.
  JOURNAL   J Virol. 1997 Feb . 71(2):891-9.
  PUBMED    8995605
REFERENCE   2  (bases 1 to 6067)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Virol. 
            1997 Feb . 71(2):891-9."
COMMENT     Sequence Label: p3187 HPV-16 E2
FEATURES             Location/Qualifiers
     source          1..6067
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(183..188,189..204,205..211)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    219..235
                     /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)."
     CDS             258..911
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     869..891
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             918..935
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(2117..2139)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2284..2302)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2370..2474
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2475..2543,2544..3335)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2693..2712)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3506..4094
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3995..4014
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4248..4265
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4338..4415
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4416..5498
                     /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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(4435..4454)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5547..5552,5553..5570,5571..5577)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5585..5601
                     /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)."
     primer_bind     5609..5625
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5609..5625
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(5621..6067,1..81)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5641..5658)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5641..5657)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5650..5672)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 agcttatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg
       61 gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt
      121 tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc
      181 tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca
      241 cacaggaaac agtattcatg tcccctatac taggttattg gaaaattaag ggccttgtgc
      301 aacccactcg acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc
      361 gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc
      421 ttccttatta tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata
      481 tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc
      541 ttgaaggagc ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact
      601 ttgaaactct caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag
      661 atcgtttatg tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt
      721 tgtatgacgc tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa
      781 aattagtttg ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat
      841 ccagcaagta tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc
      901 atcctccaaa atcggatctg gttccgcgtg gatcccccat ggagactctt tgccaacgtt
      961 taaatgtgtg tcaggacaaa atactaacac attatgaaaa tgatagtaca gacctacgtg
     1021 accatataga ctattggaaa cacatgcgcc tagaatgtgc tatttattac aaggccagag
     1081 aaatgggatt taaacatatt aaccaccagg tggtgccaac actggctgta tcaaagaata
     1141 aagcattaca agcaattgaa ctgcaactaa cgttagaaac aatatataac tcacaatata
     1201 gtaatgaaaa gtggacatta caagacgtta gccttgaagt gtatttaact gcaccaacag
     1261 gatgtataaa aaaacatgga tatacagtgg aagtgcagtt tgatggagac atatgcaata
     1321 caatgcatta tacaaactgg acacatatat atatttgtga agaagcatca gtaactgtgg
     1381 tagagggtca agttgactat tatggtttat attatgttca tgaaggaata cgaacatatt
     1441 ttgtgcagtt taaagatgat gcagaaaaat atagtaaaaa taaagtatgg gaagttcatg
     1501 cgggtggtca ggtaatatta tgtcctacat ctgtgtttag cagcaacgaa gtatcctctc
     1561 ctgaaattat taggcagcac ttggccaacc accccgccgc gacccatacc aaagccgtcg
     1621 ccttgggcac cgaagaaaca cagacgacta tccagcgacc aagatcagag ccagacaccg
     1681 gaaacccctg ccacaccact aagttgttgc acagagactc agtggacagt gctccaatcc
     1741 tcactgcatt taacagctca cacaaaggac ggattaactg taatagtaac actacaccca
     1801 tagtacattt aaaaggtgat gctaatactt taaaatgttt aagatataga tttaaaaagc
     1861 attgtacatt gtatactgca gtgtcgtcta catggcattg gacaggacat aatgtaaaac
     1921 ataaaagtgc aattgttaca cttacatatg atagtgaatg gcaacgtgac caatttttgt
     1981 ctcaagttaa aataccaaaa actattacag tgtctactgg atttatgtct atatgacatg
     2041 agatcccccg ggctgcagga attcatcgtg actgactgac gatctgcctc gcgcgtttcg
     2101 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     2161 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     2221 ggggcgcagc catgacccag tcacgtagcg atagcggagt gtataattct tgaagacgaa
     2281 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     2341 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     2401 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     2461 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     2521 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     2581 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     2641 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     2701 gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt
     2761 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     2821 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     2881 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     2941 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     3001 gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac
     3061 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     3121 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     3181 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     3241 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     3301 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     3361 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     3421 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     3481 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     3541 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     3601 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     3661 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     3721 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     3781 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     3841 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     3901 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     3961 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     4021 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     4081 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     4141 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     4201 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     4261 gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     4321 cacaccgcat aaattccgac accatcgaat ggcgcaaaac ctttcgcggt atggcatgat
     4381 agcgcccgga agagagtcaa ttcagggtgg tgaatgtgaa accagtaacg ttatacgatg
     4441 tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg cgtggtgaac caggccagcc
     4501 acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat ggcggagctg aattacattc
     4561 ccaaccgcgt ggcacaacaa ctggcgggca aacagtcgtt gctgattggc gttgccacct
     4621 ccagtctggc cctgcacgcg ccgtcgcaaa ttgtcgcggc gattaaatct cgcgccgatc
     4681 aactgggtgc cagcgtggtg gtgtcgatgg tagaacgaag cggcgtcgaa gcctgtaaag
     4741 cggcggtgca caatcttctc gcgcaacgcg tcagtgggct gatcattaac tatccgctgg
     4801 atgaccagga tgccattgct gtggaagctg cctgcactaa tgttccggcg ttatttcttg
     4861 atgtctctga ccagacaccc atcaacagta ttattttctc ccatgaagac ggtacgcgac
     4921 tgggcgtgga gcatctggtc gcattgggtc accagcaaat cgcgctgtta gcgggcccat
     4981 taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca taaatatctc actcgcaatc
     5041 aaattcagcc gatagcggaa cgggaaggcg actggagtgc catgtccggt tttcaacaaa
     5101 ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat gctggttgcc aacgatcaga
     5161 tggcgctggg cgcaatgcgc gccattaccg agtccgggct gcgcgttggt gcggatatct
     5221 cggtagtggg atacgacgat accgaagaca gctcatgtta tatcccgccg ttaaccacca
     5281 tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga ccgcttgctg caactctctc
     5341 agggccaggc ggtgaagggc aatcagctgt tgcccgtctc actggtgaaa agaaaaacca
     5401 ccctggcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     5461 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtaagt
     5521 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     5581 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacggattc
     5641 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
     5701 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
     5761 cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc tttgcctggt ttccggcacc
     5821 agaagcggtg ccggaaagct ggctggagtg cgatcttcct gaggccgata ctgtcgtcgt
     5881 cccctcaaac tggcagatgc acggttacga tgcgcccatc tacaccaacg tgacctatcc
     5941 cattacggtc aatccgccgt ttgttcccac ggagaatccg acgggttgtt actcgctcac
     6001 atttaatgtt gatgaaagct ggctacagga aggccagacg cgaattattt ttgatggcgt
     6061 tggaatt
//
