LOCUS       sequence-443105-        5244 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 5244)
  AUTHORS   Wu EW, Clemens KE, Heck DV, Munger K
  TITLE     The human papillomavirus E7 oncoprotein and the cellular 
            transcription factor E2F bind to separate sites on the 
            retinoblastoma tumor suppressor protein.
  JOURNAL   J Virol. 1993 Apr . 67(4):2402-7.
  PUBMED    8445736
REFERENCE   2  (bases 1 to 5244)
  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. 
            1993 Apr . 67(4):2402-7."
COMMENT     Sequence Label: pGEX2T E7 Q
FEATURES             Location/Qualifiers
     source          1..5244
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..6,7..22,23..29)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    37..53
                     /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             76..729
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     687..709
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             736..753
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(1112..1134)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1279..1297)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1365..1469
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1470..1538,1539..2330)
                     /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(1688..1707)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2501..3089
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2990..3009
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3243..3260
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3333..3410
                     /gene="lacI"
                     /label=lacI promoter
     CDS             3411..4493
                     /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(3430..3449)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(4542..4547,4548..4565,4566..4572)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4580..4596
                     /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     4604..4620
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4604..4620
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4616..5143
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(4636..4653)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4636..4652)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4645..4667)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 ttgacaatta atcatcggct cgtataatgt gtggaattgt gagcggataa caatttcaca
       61 caggaaacag tattcatgtc ccctatacta ggttattgga aaattaaggg ccttgtgcaa
      121 cccactcgac ttcttttgga atatcttgaa gaaaaatatg aagagcattt gtatgagcgc
      181 gatgaaggtg ataaatggcg aaacaaaaag tttgaattgg gtttggagtt tcccaatctt
      241 ccttattata ttgatggtga tgttaaatta acacagtcta tggccatcat acgttatata
      301 gctgacaagc acaacatgtt gggtggttgt ccaaaagagc gtgcagagat ttcaatgctt
      361 gaaggagcgg ttttggatat tagatacggt gtttcgagaa ttgcatatag taaagacttt
      421 gaaactctca aagttgattt tcttagcaag ctacctgaaa tgctgaaaat gttcgaagat
      481 cgtttatgtc ataaaacata tttaaatggt gatcatgtaa cccatcctga cttcatgttg
      541 tatgacgctc ttgatgttgt tttatacatg gacccaatgt gcctggatgc gttcccaaaa
      601 ttagtttgtt ttaaaaaacg tattgaagct atcccacaaa ttgataagta cttgaaatcc
      661 agcaagtata tagcatggcc tttgcagggc tggcaagcca cgtttggtgg tggcgaccat
      721 cctccaaaat cggatctggt tccgcgtgga tccaccatgc atggagatac acctacattg
      781 catgaatata tgttagattt gcaaccagag acaactgatc tctactgtta tgagcaatta
      841 aatcagagct cacagcagca gcagcaaata gatggtccag ctggacaagc agaaccggac
      901 agagcccatt acaatattgt aaccttttgt tgcaagtgtg actctacgct tcggttgtgc
      961 gtacaaagca cacacgtaga cattcgtact ttggaagacc tgttaatggg cacactagga
     1021 attgtgtgcc ccatctgttc tcagaaacca taagaattca tcgtgactga ctgacgatct
     1081 gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     1141 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     1201 gtgttggcgg gtgtcggggc gcagccatga cccagtcacg tagcgatagc ggagtgtata
     1261 attcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     1321 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     1381 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     1441 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     1501 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     1561 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     1621 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     1681 aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc
     1741 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     1801 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     1861 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     1921 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     1981 taccaaacga cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac
     2041 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     2101 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     2161 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     2221 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     2281 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     2341 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     2401 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     2461 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     2521 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     2581 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     2641 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     2701 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     2761 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     2821 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     2881 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     2941 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     3001 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     3061 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     3121 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     3181 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     3241 gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc
     3301 atctgtgcgg tatttcacac cgcataaatt ccgacaccat cgaatggcgc aaaacctttc
     3361 gcggtatggc atgatagcgc ccggaagaga gtcaattcag ggtggtgaat gtgaaaccag
     3421 taacgttata cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg
     3481 tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg
     3541 agctgaatta cattcccaac cgcgtggcac aacaactggc gggcaaacag tcgttgctga
     3601 ttggcgttgc cacctccagt ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta
     3661 aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg
     3721 tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca
     3781 ttaactatcc gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc
     3841 cggcgttatt tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg
     3901 aagacggtac gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc
     3961 tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat
     4021 atctcactcg caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt
     4081 ccggttttca acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg
     4141 ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg
     4201 ttggtgcgga tatctcggta gtgggatacg acgataccga agacagctca tgttatatcc
     4261 cgccgttaac caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct
     4321 tgctgcaact ctctcagggc caggcggtga agggcaatca gctgttgccc gtctcactgg
     4381 tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     4441 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     4501 gcaattaatg taagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
     4561 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
     4621 catgattacg gattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     4681 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     4741 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgctttgc
     4801 ctggtttccg gcaccagaag cggtgccgga aagctggctg gagtgcgatc ttcctgaggc
     4861 cgatactgtc gtcgtcccct caaactggca gatgcacggt tacgatgcgc ccatctacac
     4921 caacgtgacc tatcccatta cggtcaatcc gccgtttgtt cccacggaga atccgacggg
     4981 ttgttactcg ctcacattta atgttgatga aagctggcta caggaaggcc agacgcgaat
     5041 tatttttgat ggcgttggaa ttagcttatc gactgcacgg tgcaccaatg cttctggcgt
     5101 caggcagcca tcggaagctg tggtatggct gtgcaggtcg taaatcactg cataattcgt
     5161 gtcgctcaag gcgcactccc gttctggata atgttttttg cgccgacatc ataacggttc
     5221 tggcaaatat tctgaaatga gctg
//
