LOCUS       sequence-175722-        7462 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 7462)
  AUTHORS   Talis AL, Huibregtse JM, Howley PM
  TITLE     The role of E6AP in the regulation of p53 protein levels in human 
            papillomavirus (HPV)-positive and HPV-negative cells.
  JOURNAL   J Biol Chem 1998 Mar 13;273(11):6439-45.
  PUBMED    9497376
REFERENCE   2  (bases 1 to 7462)
  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 Biol 
            Chem"; date: "1998-03-13"; volume: "273(11)"; pages: "6439-45"
COMMENT     Sequence Label: p3868 HA-E6-AP
FEATURES             Location/Qualifiers
     source          1..7462
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     59..81
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     73..90
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     74..90
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        251..630
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        631..834
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     785..805
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             911..937
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    3563..4185
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3762..3781)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(4209..4229)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4214..4543
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4394..4529
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          4582..4678
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     primer_bind     complement(4754..4773)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4755..4773)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4787..4803)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4787..4803)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4800..4822)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4811..4827
                     /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(4835..4841,4842..4859,4860..4865))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4880..4901
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5018..5035)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5189..5777)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5269..5288)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5948..6739,6740..6808))
                     /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     6571..6590
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6809..6913)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6940..7395
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7027..7046)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7237..7258
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
       61 cagtcacgac gttgtaaaac gacggccagt gccaagctga tctatacatt gaatcaatat
      121 tggcaattag ccatattagt cattggttat atagcataaa tcaatattgg ctattggcca
      181 ttgcatacgt tgtatctata tcataatatg tacatttata ttggctcatg tccaatatga
      241 ccgccatgtt gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
      301 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
      361 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      421 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      481 gcagtacatc aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa
      541 tggcccgcct ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac
      601 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg
      661 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
      721 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg
      781 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta
      841 gtgaaccgtc agaattgttt ttatttttaa ttttctttca aatacttcca tcgaattcag
      901 atccaccatg tacccatacg atgttccaga ttacgctgaa gcctgcacga atgagttttg
      961 tgcttcctgt ccaacttttc ttcgtatgga taataatgca gcagctatta aagccctcga
     1021 gctttataag attaatgcaa aactctgtga tcctcatccc tccaagaaag gagcaagctc
     1081 agcttacctt gagaactcga aaggtgcccc caacaactcc tgctctgaga taaaaatgaa
     1141 caagaaaggc gctagaattg attttaaaga tgtgacttac ttaacagaag agaaggtata
     1201 tgaaattctt gaattatgta gagaaagaga ggattattcc cctttaatcc gtgttattgg
     1261 aagagttttt tctagtgctg aggcattggt acagagcttc cggaaagtta aacaacacac
     1321 caaggaagaa ctgaaatctc ttcaagcaaa agatgaagac aaagatgaag atgaaaagga
     1381 aaaagctgca tgttctgctg ctgctatgga agaagactca gaagcatctt cctcaaggat
     1441 aggtgatagc tcacagggag acaacaattt gcaaaaatta ggccctgatg atgtgtctgt
     1501 ggatattgat gccattagaa gggtctacac cagattgctc tctaatgaaa aaattgaaac
     1561 tgcctttctc aatgcacttg tatatttgtc acctaacgtg gaatgtgact tgacgtatca
     1621 caatgtatac tctcgagatc ctaattatct gaatttgttc attatcgtaa tggagaatag
     1681 aaatctccac agtcctgaat atctggaaat ggctttgcca ttattttgca aagcgatgag
     1741 caagctaccc cttgcagccc aaggaaaact gatcagactg tggtctaaat acaatgcaga
     1801 ccagattcgg agaatgatgg agacatttca gcaacttatt acttataaag tcataagcaa
     1861 tgaatttaac agtcgaaatc tagtgaatga tgatgatgcc attgttgctg cttcgaagtg
     1921 cttgaaaatg gtttactatg caaatgtagt gggaggggaa gtggacacaa atcacaatga
     1981 agaagatgat gaagagccca tccctgagtc cagcgagctg acacttcagg aacttttggg
     2041 agaagaaaga agaaacaaga aaggtcctcg agtggacccc ctggaaactg aacttggtgt
     2101 taaaaccctg gattgtcgaa aaccacttat cccttttgaa gagtttatta atgaaccact
     2161 gaatgaggtt ctagaaatgg ataaagatta tacttttttc aaagtagaaa cagagaacaa
     2221 attctctttt atgacatgtc cctttatatt gaatgctgtc acaaagaatt tgggattata
     2281 ttatgacaat agaattcgca tgtacagtga acgaagaatc actgttctct acagcttagt
     2341 tcaaggacag cagttgaatc catatttgag actcaaagtt agacgtgacc atatcataga
     2401 tgatgcactt gtccggctag agatgatcgc tatggaaaat cctgcagact tgaagaagca
     2461 gttgtatgtg gaatttgaag gagaacaagg agttgatgag ggaggtgttt ccaaagaatt
     2521 ttttcagctg gttgtggagg aaatcttcaa tccagatatt ggtatgttca catacgatga
     2581 atctacaaaa ttgttttggt ttaatccatc ttcttttgaa actgagggtc agtttactct
     2641 gattggcata gtactgggtc tggctattta caataactgt atactggatg tacattttcc
     2701 catggttgtc tacaggaagc taatggggaa aaaaggaact tttcgtgact tgggagactc
     2761 tcacccagtt ctatatcaga gtttaaaaga tttattggag tatgaaggga atgtggaaga
     2821 tgacatgatg atcactttcc agatatcaca gacagatctt tttggtaacc caatgatgta
     2881 tgatctaaag gaaaatggtg ataaaattcc aattacaaat gaaaacagga aggaatttgt
     2941 caatctttat tctgactaca ttctcaataa atcagtagaa aaacagttca aggcttttcg
     3001 gagaggtttt catatggtga ccaatgaatc tcccttaaag tacttattca gaccagaaga
     3061 aattgaattg cttatatgtg gaagccggaa tctagatttc caagcactag aagaaactac
     3121 agaatatgac ggtggctata ccagggactc tgttctgatt agggagttct gggaaatcgt
     3181 tcattcattt acagatgaac agaaaagact cttcttgcag tttacaacgg gcacagacag
     3241 agcacctgtg ggaggactag gaaaattaaa gatgattata gccaaaaatg gcccagacac
     3301 agaaaggtta cctacatctc atacttgctt taatgtgctt ttacttccgg aatactcaag
     3361 caaagaaaaa cttaaagaga gattgttgaa ggccatcacg tatgccaaag gatttggcat
     3421 gctgtaaaac gcggccgctg cagatctgat cctttcctgg gacccggcaa gaaccaaaaa
     3481 ctcactctct tcaaggaaat ccgtaatgtt aaacccgaca cgatgaagct tgcatgcctg
     3541 caggtcgact ctagaggatc ccgggtggca tccctgtgac ccctccccag tgcctctcct
     3601 ggccctggaa gttgccactc cagtgcccac cagccttgtc ctaataaaat taagttgcat
     3661 cattttgtct gactaggtgt ccttctataa tattatgggg tggagggggg tggtatggag
     3721 caaggggcaa gttgggaaga caacctgtag ggcctgcggg gtctattggg aaccaagctg
     3781 gagtgcagtg gcacaatctt ggctcactgc aatctccgcc tcctgggttc aagcgattct
     3841 cctgcctcag cctcccgagt tgttgggatt ccaggcatgc atgaccaggc tcagctaatt
     3901 tttgtttttt tggtagagac ggggtttcac catattggcc aggctggtct ccaactccta
     3961 atctcaggtg atctacccac cttggcctcc caaattgctg ggattacagg cgtgaaccac
     4021 tgctcccttc cctgtccttc tgattttaaa ataactatac cagcaggagg acgtccagac
     4081 acagcatagg ctacctggcc atgcccaacc ggtgggacat ttgagttgct tgcttggcac
     4141 tgtcctctca tgcgttgggt ccactcagta gatgcctgtt gaattgggta cgcggccagc
     4201 ttggctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     4261 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     4321 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     4381 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     4441 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     4501 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc tcgaggaact
     4561 gaaaaaccag aaagttaact ggtaagttta gtctttttgt cttttatttc aggtcccgga
     4621 tccggtggtg gtgcaaatca aagaactgct cctcagtgga tgttgccttt acttctaggc
     4681 ctgtacggaa gtgttacttc tgctctaaaa gctgctgcag gtcgactcta gggccgcgta
     4741 ccgagctcga attccctata gtgagtcgta ttaaattcgt aatcatgtca tagctgtttc
     4801 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     4861 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     4921 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     4981 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     5041 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     5101 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     5161 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     5221 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     5281 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     5341 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     5401 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     5461 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     5521 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     5581 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     5641 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     5701 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     5761 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5821 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5881 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     5941 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     6001 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     6061 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     6121 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     6181 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     6241 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     6301 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     6361 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6421 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6481 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     6541 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     6601 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6661 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6721 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6781 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6841 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6901 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgcgccctgt agcggcgcat
     6961 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     7021 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     7081 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     7141 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     7201 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     7261 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     7321 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     7381 taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     7441 gcgggcctct tcgctattac gc
//
