LOCUS       V023992                 8335 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023992
VERSION     V023992
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     misc_signal     190..340
                     /note="packaging signal for adenovirus serotype 5"
                     /label="Ad5 Ψ"
     promoter        470..993
                     /label="mCMV promoter"
     CDS             1006..5364
                     /label="Col1a1(NM_007742)"
                     /note="Col1a1(NM_007742)"
                     /gene="Col1a1"
     CDS             5371..5442
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags"
                     /transl_table=1
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    5458..5592
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     protein_bind    complement(5612..5645)
                     /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(5667..5683)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5691..5707
                     /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(5715..5745)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5760..5781
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6069..6657)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6828..7688)
                     /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(7689..7793)
                     /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 cagctctaga 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 atttaattaa aacgccatgt actttcccac
      721 cattgacgtc aatgggctat tgaaactaat gcaacgtgac ctttaaacgg tactttccca
      781 tagctgatta atgggaaagt accgttctcg agccaataca cgtcaatggg aagtgaaagg
      841 gcagccaaaa cgtaacaccg ccccggtttt cccctggaaa ttccatattg gcacgcattc
      901 tattggctga gctgcgttct acgtgggtat aagaggcgcg accagcgtcg gtaccgtcgc
      961 agtcttcggt ctgaccaccg tagaacgcag atcgaattcg ccaccatgtt cagctttgtg
     1021 gacctccggc tcctgctcct cttaggggcc actgccctcc tgacgcatgg ccaagaagac
     1081 atccctgaag tcagctgcat acacaatggc ctaagggtcc ccaatggtga gacgtggaaa
     1141 cccgaggtat gcttgatctg tatctgccac aatggcacgg ctgtgtgcga tgacgtgcaa
     1201 tgcaatgaag aactggactg tcccaacccc caaagacggg agggcgagtg ctgtgctttc
     1261 tgcccggaag aatacgtatc accaaactca gaagatgtag gagtcgaggg acccaaggga
     1321 gaccctggcc cccaaggccc aaggggaccc gttggccccc ctggacgaga tggcatccct
     1381 ggacagcctg gacttcctgg tcctcctggt ccccctgggc cccccggacc ccctggcctt
     1441 ggaggaaact ttgcttccca gatgtcctat ggctatgatg aaaaatcagc tggagtttcc
     1501 gtgcctggcc ccatgggtcc ttctggtcct cgtggtctcc ctggcccccc tggtgcacct
     1561 ggtccacaag gtttccaagg cccccctggt gaacctggcg agcctggcgg ttcaggtcca
     1621 atgggtcccc gaggtccccc tggccctcct ggcaagaatg gagatgatgg ggaagctggc
     1681 aagcccggcc gtcctggtga gcgtggacct cctggacctc agggtgctcg tggattgcct
     1741 ggaacagctg gcctccctgg aatgaaggga caccgaggct tcagtggttt ggatggtgcc
     1801 aaaggagatg ctggtcctgc tggtcctaag ggagagcccg gcagtcctgg tgaaaacgga
     1861 gctcctggcc agatgggtcc ccgaggtctg cccggtgaga gaggtcgccc tggacctcct
     1921 ggcactgctg gtgctcgcgg taacgatggt gctgttggtg ctgctggacc ccctggtccc
     1981 accggcccca ctggccctcc tggcttccct ggtgcagttg gtgctaaggg tgaagctggt
     2041 ccccaaggag ctagaggctc tgaaggtccc cagggtgtgc gtggtgagcc cggaccccct
     2101 ggccctgctg gtgctgccgg ccctgctgga aaccctggtg ctgatggaca acctggcgct
     2161 aaaggtgcca atggtgctcc tggtattgct ggtgctcctg gcttccctgg tgcccgaggc
     2221 ccctctggac cccagggccc cagcggccct ccaggtccca agggtaacag tggtgaacct
     2281 ggtgctcctg gcaacaaagg agacactggt gccaaaggag aacccggtgc tactggagtt
     2341 caaggtcccc caggccctgc cggagaagaa ggaaaacgag gagcccgtgg tgagcctgga
     2401 ccttccggac tgcctggacc tcctggcgag cgtggtggac ctggtagccg tggtttccct
     2461 ggtgctgatg gtgttgctgg ccccaagggt ccttccggtg aacgtggtgc tcccggacct
     2521 gctggtccca aaggttctcc tggtgaagct ggtcgccccg gtgaagctgg tctccctggt
     2581 gccaagggtc tcactggcag tcctggcagc cctggtcctg atggcaaaac cggcccccct
     2641 ggtcccgctg gtcaagatgg tcgccctgga cccgcaggtc ctcctggagc ccgtggccag
     2701 gctggtgtga tgggattccc tggacctaag ggtaccgctg gagaacctgg aaaggctgga
     2761 gagcgaggcc ttcccggacc ccctggcgct gttggtcctg ctggcaaaga tggagaagct
     2821 ggagctcagg gagcccctgg ccctgctggt cctgctggtg agagaggtga acaaggtccc
     2881 gctggctccc ctggattcca gggtcttcct ggtcctgccg gtcctcctgg tgaagcaggc
     2941 aagcctggtg aacagggtgt tcctggagac cttggtgccc ctggaccctc tggcgcaaga
     3001 ggcgagagag gtttccctgg tgaacgtggt gtacaaggtc ccccaggtcc tgctggtccc
     3061 cgaggaaaca atggtgcccc cggcaacgat ggtgccaagg gtgatactgg tgcccccgga
     3121 gctcccggta gccagggtgc ccccggtctt cagggaatgc ctggtgaacg tggtgcagct
     3181 ggtcttccag gtcctaaggg tgacagaggt gatgctggtc ccaaaggtgc tgatggttct
     3241 cctggtaaag atggtgcccg tggtctgact ggtcccattg gtcctcctgg ccctgctggt
     3301 gcccctggtg acaagggtga agctggtccc agtggtcctc ccggtcccac cggagcccgt
     3361 ggtgctcccg gagaccgtgg tgaggctggt ccccctggtc ctgctggctt tgccggcccc
     3421 cctggtgctg atggccaacc tggtgcgaaa ggtgaacctg gtgatactgg tgttaaaggt
     3481 gatgctggtc ctcctggccc tgctggtcct gctggacccc ccggccccat tggtaacgtt
     3541 ggtgctcctg gacccaaagg tcctcgtggt gctgctggtc cccctggtgc tactggcttc
     3601 cctggtgctg ctggccgtgt cggtccccct ggtccctctg gaaatgctgg accccctggc
     3661 cctcccggtc ccgttggcaa agaagggggc aaaggtcccc gtggtgagac tggccctgct
     3721 ggacgtcctg gtgaagttgg tcccccaggt ccccccggtc ctgctggtga gaaaggatct
     3781 cctggtgctg atggacctgc tggctctcct ggtacccctg gacctcaggg tattgctgga
     3841 caacgtggtg tggtcggtct tcccggtcag agaggagaaa gaggcttccc tggtcttcct
     3901 ggcccctctg gtgaacctgg caaacaaggt ccttctggat caagtggtga acgcggtccc
     3961 cctggcccca tggggccccc tggattggct ggtccccctg gtgaatctgg acgtgaggga
     4021 tcccctggtg ctgaaggctc ccctggaagg gatggtgctc ccggggccaa gggtgaccgt
     4081 ggtgagactg gccccgctgg cccccctggt gcccctggtg ctcccggtgc tcccggccct
     4141 gttggtcccg ctggcaagaa tggcgatcgt ggtgagactg gtcctgctgg tcctgctggt
     4201 cccattggcc ctgctggtgc ccgtggccct gctggacccc aaggcccccg tggtgacaag
     4261 ggtgagacag gcgaacaagg tgacagaggc ataaagggtc atcgtggctt ctctggtctc
     4321 cagggtcctc ctggttctcc tggttctcct ggtgaacaag gcccctctgg agcttcaggt
     4381 cctgcaggcc cccggggtcc ccctggctct gctggttctc ctggcaaaga cggactcaac
     4441 ggtctccctg gccccattgg tccccctggt cctcgaggtc gcactggtga cagcggccct
     4501 gctggtcccc ccggccctcc tggaccccct ggccctcctg gacctcccag tggcggttat
     4561 gacttcagct tcctgcctca gccacctcaa gagaagtctc aagatggtgg ccgctactac
     4621 cgggccgatg atgctaacgt ggttcgtgac cgtgaccttg aggtggacac caccctcaag
     4681 agcctgagtc agcagattga gaacatccgc agccccgaag gcagccgcaa gaaccctgcc
     4741 cgcacatgcc gcgacctcaa gatgtgccac tctgactgga agagcggaga gtactggatc
     4801 gaccctaacc aaggctgcaa cctggacgcc atcaaggtct actgcaacat ggagacaggt
     4861 cagacctgtg tgttccctac tcagccgtct gtgcctcaga agaactggta catcagcccg
     4921 aaccccaagg aaaagaagca cgtctggttt ggagagagca tgaccgatgg attcccgttc
     4981 gagtacggaa gcgagggctc cgaccccgcc gatgtcgcta tccagctgac cttcctgcgc
     5041 ctaatgtcca ccgaggcctc ccagaacatc acctatcact gcaagaacag cgtagcctac
     5101 atggaccagc agactggcaa cctcaagaag gccctgctcc tccagggatc caacgagatc
     5161 gagctcagag gcgaaggcaa cagtcgcttc acctacagca cccttgtgga cggctgcacg
     5221 agtcacaccg gaacttgggg caagacagtc atcgaataca aaaccaccaa gacctcccgc
     5281 ctgcccatca tcgatgtggc tcccttggac attggtgccc cagaccagga attcggacta
     5341 gacattggcc ctgcctgctt cgtgaagctt gattacaagg atgacgacga taaggactat
     5401 aaggacgatg atgacaagga ctacaaagat gatgacgata aatagtcgac ttcgagcaac
     5461 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     5521 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     5581 catgtctgga tcgtctagca tcgaagatcc aataacttcg tatagcatac attatacgaa
     5641 gttataagta gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     5701 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     5761 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     5821 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     5881 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     5941 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     6001 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     6061 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     6121 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     6181 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     6241 tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc
     6301 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     6361 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     6421 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     6481 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     6541 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     6601 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     6661 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     6721 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     6781 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     6841 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     6901 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     6961 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     7021 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     7081 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     7141 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     7201 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     7261 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     7321 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     7381 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     7441 tcaacacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     7501 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     7561 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     7621 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     7681 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     7741 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     7801 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
     7861 aataggcgta tcactctagg caaaatagca ccctcccgct ccagaacaac atacagcgct
     7921 tcacagcggc agcctaacag tcagccttac cagtaaaaaa gaaaacctat taaaaaaaca
     7981 ccactcgaca cggcaccagc tcaatcagtc acagtgtaaa aaagggccaa gtgcagagcg
     8041 agtatatata ggactaaaaa atgacgtaac ggttaaagtc cacaaaaaac acccagaaaa
     8101 ccgcacgcga acctacgccc agaaacgaaa gccaaaaaac ccacaacttc ctcaaatcgt
     8161 cacttccgtt ttcccacgtt acgtaacttc ccattttaag aaaactacaa ttcccaacac
     8221 atacaagtta ctccgcccta aaacctacgt cacccgcccc gttcccacgc cccgcgccac
     8281 gtcacaaact ccaccccctc attatcatat tggcttcaat ccaaaataag gtata
//
