LOCUS       sequence-436568-        5969 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 5969)
  AUTHORS   Sakai H, Yasugi T, Benson JD, Dowhanick JJ, Howley PM
  TITLE     Targeted mutagenesis of the human papillomavirus type 16 E2 
            transactivation domain reveals separable transcriptional activation 
            and DNA replication functions.
  JOURNAL   J Virol. 1996 Mar . 70(3):1602-11.
  PUBMED    8627680
REFERENCE   2  (bases 1 to 5969)
  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. 
            1996 Mar . 70(3):1602-11."
COMMENT     Sequence Label: p3662 HPV-16 E2
FEATURES             Location/Qualifiers
     source          1..5969
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        42..421
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        422..625
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     576..596
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      744..753
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1861..2483
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2060..2079)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2507..2527)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2512..2841
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2692..2827
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          2880..2976
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     primer_bind     complement(3052..3071)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3053..3071)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3085..3101)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3085..3101)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3098..3120)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3109..3125
                     /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)."
     promoter        complement(join(3133..3139,3140..3157,3158..3163))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3178..3199
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3316..3333)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3487..4075)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3567..3586)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4246..5037,5038..5106))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4869..4888
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5107..5211)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5238..5693
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5325..5344)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5535..5556
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5819..5841
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5833..5850
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5834..5850
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gtacatttat attggctcat gtccaatatg accgccatgt tgacattgat tattgactag
       61 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
      121 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
      181 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
      241 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
      301 tccgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
      361 gaccttacgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
      421 ggtgatgcgg ttttggcagt acaccaatgg gcgtggatag cggtttgact cacggggatt
      481 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
      541 ctttccaaaa tgtcgtaata accccgcccc gttgacgcaa atgggcggta ggcgtgtacg
      601 gtgggaggtc tatataagca gagctcgttt agtgaaccgt cagaattgtt tttattttta
      661 attttctttc aaatacttcc atcgaattca gatctggtac cacgcgtatc gataagcttg
      721 catgcctgca ggtcgactct agagccacca tggagactct ttgccaacgt ttaaatgtgt
      781 gtcaggacaa aatactaaca cattatgaaa atgatagtac agacctacgt gaccatatag
      841 actattggaa acacatgcgc ctagaatgtg ctatttatta caaggccaga gaaatgggat
      901 ttaaacatat taaccaccag gtggtgccaa cactggctgt atcaaagaat aaagcattac
      961 aagcaattga actgcaacta acgttagaaa caatatataa ctcacaatat agtaatgaaa
     1021 agtggacatt acaagacgtt agccttgaag tgtatttaac tgcaccaaca ggatgtataa
     1081 aaaaacatgg atatacagtg gaagtgcagt ttgatggaga catatgcaat acaatgcatt
     1141 atacaaactg gacacatata tatatttgtg aagaagcatc agtaactgtg gtagagggtc
     1201 aagttgacta ttatggttta tattatgttc atgaaggaat acgaacatat tttgtgcagt
     1261 ttaaagatga tgcagaaaaa tatagtaaaa ataaagtatg ggaagttcat gcgggtggtc
     1321 aggtaatatt atgtcctaca tctgtgttta gcagcaacga agtatcctct cctgaaatta
     1381 ttaggcagca cttggccaac caccccgccg cgacccatac caaagccgtc gccttgggca
     1441 ccgaagaaac acagacgact atccagcgac caagatcaga gccagacacc ggaaacccct
     1501 gccacaccac taagttgttg cacagagact cagtggacag tgctccaatc ctcactgcat
     1561 ttaacagctc acacaaagga cggattaact gtaatagtaa cactacaccc atagtacatt
     1621 taaaaggtga tgctaatact ttaaaatgtt taagatatag atttaaaaag cattgtacat
     1681 tgtatactgc agtgtcgtct acatggcatt ggacaggaca taatgtaaaa cataaaagtg
     1741 caattgttac acttacatat gatagtgaat ggcaacgtga ccaatttttg tctcaagtta
     1801 aaataccaaa aactattaca gtgtctactg gatttatgtc tatatgacat gagatccccc
     1861 gggtggcatc cctgtgaccc ctccccagtg cctctcctgg ccctggaagt tgccactcca
     1921 gtgcccacca gccttgtcct aataaaatta agttgcatca ttttgtctga ctaggtgtcc
     1981 ttctataata ttatggggtg gaggggggtg gtatggagca aggggcaagt tgggaagaca
     2041 acctgtaggg cctgcggggt ctattgggaa ccaagctgga gtgcagtggc acaatcttgg
     2101 ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg
     2161 ttgggattcc aggcatgcat gaccaggctc agctaatttt tgtttttttg gtagagacgg
     2221 ggtttcacca tattggccag gctggtctcc aactcctaat ctcaggtgat ctacccacct
     2281 tggcctccca aattgctggg attacaggcg tgaaccactg ctcccttccc tgtccttctg
     2341 attttaaaat aactatacca gcaggaggac gtccagacac agcataggct acctggccat
     2401 gcccaaccgg tgggacattt gagttgcttg cttggcactg tcctctcatg cgttgggtcc
     2461 actcagtaga tgcctgttga attgggtacg cggccagctt ggctgtggaa tgtgtgtcag
     2521 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     2581 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     2641 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     2701 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     2761 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     2821 ggaggcctag gcttttgcaa aaagctcctc gaggaactga aaaaccagaa agttaactgg
     2881 taagtttagt ctttttgtct tttatttcag gtcccggatc cggtggtggt gcaaatcaaa
     2941 gaactgctcc tcagtggatg ttgcctttac ttctaggcct gtacggaagt gttacttctg
     3001 ctctaaaagc tgctgcaggt cgactctagg gccgcgtacc gagctcgaat tccctatagt
     3061 gagtcgtatt aaattcgtaa tcatgtcata gctgtttcct gtgtgaaatt gttatccgct
     3121 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     3181 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     3241 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     3301 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     3361 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     3421 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     3481 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     3541 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     3601 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     3661 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     3721 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     3781 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     3841 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     3901 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     3961 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     4021 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     4081 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     4141 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     4201 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     4261 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     4321 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     4381 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     4441 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     4501 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     4561 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     4621 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     4681 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     4741 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     4801 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     4861 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     4921 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     4981 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     5041 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     5101 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     5161 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     5221 ccgaaaagtg ccacctgacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     5281 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     5341 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     5401 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     5461 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     5521 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     5581 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     5641 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttccattcg
     5701 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     5761 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
     5821 cagtcacgac gttgtaaaac gacggccagt gccaagctga tctatacatt gaatcaatat
     5881 tggcaattag ccatattagt cattggttat atagcataaa tcaatattgg ctattggcca
     5941 ttgcatacgt tgtatctata tcataatat
//
