LOCUS       V032986                 5851 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032986
VERSION     V032986
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     misc_signal     190..340
                     /note="packaging signal for adenovirus serotype 5"
                     /label="Ad5 Ψ"
     promoter        470..992
                     /label="mCMV promoter"
     CDS             1005..2915
                     /label="GSPT1(NM_002094)"
                     /note="GSPT1(NM_002094)"
                     /gene="GSPT1"
     CDS             2916..2933
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     polyA_signal    2974..3108
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     protein_bind    complement(3128..3161)
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (ATGTATGC) (Shaw et al., 2021)."
                     /label="loxP"
     primer_bind     complement(3183..3199)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    3207..3223
                     /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-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(3231..3261)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    3276..3297
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(3585..4173)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(4344..5204)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(5205..5309)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 ttcatcaata atatacctta ttttggattg aagccaatat gataatgagg gggtggagtt
       61 tgtgacgtgg cgcggggcgt gggaacgggg cgggtgacgt agtagtgtgg cggaagtgtg
      121 atgttgcaag tgtggcggaa cacatgtaag cgacggatgt ggcaaaagtg acgtttttgg
      181 tgtgcgccgg tgtacacagg aagtgacaat tttcgcgcgg ttttaggcgg atgttgtagt
      241 aaatttgggc gtaaccgagt aagatttggc cattttcgcg ggaaaactga ataagaggaa
      301 gtgaaatctg aataattttg tgttactcat agcgcgtaat atttgtctag ggccgcgggg
      361 actttgaccg tttacgtgga gactcgccca ggtgtttttc tcaggtgttt tccgcgttcc
      421 gggtcaaagt tggcgtttta ttattatagt cagntctaga gatatactga gtcattaggg
      481 actttccaat gggttttgcc cagtacataa ggtcaatagg ggtgaatcaa caggaaagtc
      541 ccattggagc caagtacact gagtcaatag ggactttcca ttgggttttg cccagtacaa
      601 aaggtcaata gggggtgagt caatgggttt ttcccattat tggcacgtac ataaggtcaa
      661 taggggtgag tcattgggtt tttccagcca tttaattaaa acgccatgta ctttcccacc
      721 attgacgtca atgggctatt gaaactaatg caacgtgacc tttaaacggt actttcccat
      781 agctgattaa tgggaaagta ccgttctcga gccaatacac gtcaatggga agtgaaaggg
      841 cagccaaaac gtaacaccgc cccggttttc ccctggaaat tccatattgg cactcattct
      901 attggctgag ctgcgttcta cgtgggtata agaggcgcga ccagcgtcgg taccgtcgca
      961 gtcttcggtc tgaccaccgt agaacgcaga tcgaattcgc caccatggat ccgggcagtg
     1021 gcggcggcgg cggcggcggc ggcggcggcg ggagcagcag cggcagcagc agcagcgact
     1081 cggcgcctga ctgctgggac caggcggaca tggaagcccc cgggccgggc ccttgcggcg
     1141 gcggcggctc cctggcggcg gcggccgagg cccagcggga gaacctcagc gcggccttca
     1201 gccggcaact caacgtcaac gccaagccct tcgtgcccaa cgtccacgcc gccgagttcg
     1261 tgccgtcctt cctgcggggc ccggcagcgc cgccaccccc agttggcggc gccgccaata
     1321 accacggagc cggcagcggc gcgggaggcc gtgcggcacc tgtggaatcc tctcaagagg
     1381 aacagtcatt gtgtgaaggt tcaaattcag ctgttagcat ggaactttca gaacctattg
     1441 tagaaaatgg agagacagaa atgtctccag aagaatcatg ggagcacaaa gaagaaataa
     1501 gtgaagcaga gccagggggt ggttccttgg gagatggaag gccgccagag gaaagtgccc
     1561 atgaaatgat ggaggaggaa gaggaaatcc caaaacctaa gtctgtggtt gcaccgccag
     1621 gtgctcctaa gaaagagcat gtaaatgtag tattcattgg gcacgtagat gctggcaagt
     1681 caaccattgg aggacaaata atgtatttga ctggaatggt tgacaaaagg acgcttgaaa
     1741 agtatgaaag agaagctaaa gagaaaaaca gagaaacttg gtacttgtct tgggccttag
     1801 acacaaatca ggaagaacga gacaagggta aaacagtaga agtgggtcgt gcctattttg
     1861 aaaccgaaaa gaagcatttc acaattctag atgcccctgg ccacaagagt tttgtcccaa
     1921 atatgattgg tggtgcctct caagctgatt tggctgtgct ggtaatctca gccaggaaag
     1981 gagagtttga aactggattt gaaaaaggag gacagacaag agaacatgca atgttggcaa
     2041 agacagcagg tgtaaaacac ctaattgtgc taattaataa gatggatgat ccaacagtaa
     2101 attggagcaa tgagagatat gaagaatgta aggagaaact agtgccattt ttgaaaaaag
     2161 ttggcttcaa tcccaaaaag gacattcact ttatgccctg ctcaggactt actggagcaa
     2221 atctcaaaga gcagtcggat ttctgtcctt ggtacattgg attaccgttt attccatatc
     2281 tggataattt gccgaacttc aatagatcag ttgatggacc aatcaggctg ccaattgtgg
     2341 ataagtacaa ggatatgggc actgtggtcc tgggaaagct ggaatcagga tctatttgta
     2401 aaggccagca gcttgtgatg atgccaaaca agcacaacgt ggaagttctt ggaatacttt
     2461 ccgatgatgt agagactgat accgtagccc caggtgaaaa cctcaaaatc agactgaaag
     2521 gaattgaaga agaggagatt cttccagggt ttatactttg tgatcctaat aatctttgtc
     2581 attctggacg cacatttgat gcccagatag tgattataga gcacaaatcc atcatctgcc
     2641 caggctataa tgcggtgctg catattcata cctgtattga ggaggtggaa ataacagcct
     2701 taatctgctt ggtagacaaa aaatcaggag aaaaaagtaa gacccgaccc cgttttgtca
     2761 aacaagatca agtatgcatt gctcgcttaa ggacagcagg aaccatctgc cttgagacct
     2821 ttaaagactt ccctcagatg ggtcgtttca ccttaagaga tgagggtaag accattgcaa
     2881 ttggaaaagt tctgaaactg gttccagaga aagaccacca ccaccaccac cactaacccg
     2941 ggagatctaa gcttgagctc gtcgacttcg agcaacttgt ttattgcagc ttataatggt
     3001 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     3061 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatcgt ctagcatcga
     3121 agatccaata acttcgtata gcatacatta tacgaagtta taagtagctt ggcgtaatca
     3181 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     3241 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     3301 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     3361 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     3421 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     3481 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     3541 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     3601 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     3661 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     3721 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa
     3781 tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     3841 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     3901 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     3961 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     4021 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     4081 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     4141 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     4201 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     4261 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     4321 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     4381 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     4441 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     4501 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     4561 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     4621 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     4681 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     4741 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     4801 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     4861 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     4921 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca
     4981 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     5041 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     5101 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     5161 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     5221 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     5281 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     5341 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac tctaggcaaa
     5401 atagcaccct cccgctccag aacaacatac agcgcttcac agcggcagcc taacagtcag
     5461 ccttaccagt aaaaaagaaa acctattaaa aaaacaccac tcgacacggc accagctcaa
     5521 tcagtcacag tgtaaaaaag ggccaagtgc agagcgagta tatataggac taaaaaatga
     5581 cgtaacggtt aaagtccaca aaaaacaccc agaaaaccgc acgcgaacct acgcccagaa
     5641 acgaaagcca aaaaacccac aacttcctca aatcgtcact tccgttttcc cacgttacgt
     5701 aacttcccat tttaagaaaa ctacaattcc caacacatac aagttactcc gccctaaaac
     5761 ctacgtcacc cgccccgttc ccacgccccg cgccacgtca caaactccac cccctcatta
     5821 tcatattggc ttcaatccaa aataaggtat a
//
