LOCUS       V045701                 7283 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045701
VERSION     V045701
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     misc_signal     190..340
                     /note="packaging signal for adenovirus serotype 5"
                     /label="Ad5 Ψ"
     promoter        469..992
                     /label="mCMV promoter"
     CDS             999..1715
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK"
                     /label="EGFP"
     polyA_signal    1862..1983
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     enhancer        1991..2370
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        2371..2574
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        2619..2637
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             2694..4061
                     /label="Foxd1(NM_008242)"
                     /note="Foxd1(NM_008242)"
                     /gene="Foxd1"
     CDS             4068..4133
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    4168..4392
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     polyA_signal    4406..4540
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     protein_bind    complement(4560..4593)
                     /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(4615..4631)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4639..4655
                     /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(4663..4693)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4708..4729
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5017..5605)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5776..6636)
                     /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(6637..6741)
                     /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 cagtctagag atatactgag tcattaggga
      481 ctttccaatg ggttttgccc agtacataag gtcaataggg gtgaatcaac aggaaagtcc
      541 cattggagcc aagtacactg agtcaatagg gactttccat tgggttttgc ccagtacaaa
      601 aggtcaatag ggggtgagtc aatgggtttt tcccattatt ggcacgtaca taaggtcaat
      661 aggggtgagt cattgggttt ttccagccaa tttaattaaa acgccatgta ctttcccacc
      721 attgacgtca atgggctatt gaaactaatg caacgtgacc tttaaacggt actttcccat
      781 agctgattaa tgggaaagta ccgttctcga gccaatacac gtcaatggga agtgaaaggg
      841 cagccaaaac gtaacaccgc cccggttttc ccctggaaat tccatattgg cacgcattct
      901 attggctgag ctgcgttcta cgtgggtata agaggcgcga ccagcgtcgg taccgtcgca
      961 gtcttcggtc tgaccaccgt agaacgcaga tcgaattcat ggtgagcaag ggcgaggagc
     1021 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     1081 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
     1141 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
     1201 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     1261 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
     1321 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     1381 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     1441 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga
     1501 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     1561 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc
     1621 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     1681 ccgggatcac tctcggcatg gacgagctgt acaagtccgg actcagatct cgagatccac
     1741 cggatctaga taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt
     1801 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     1861 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     1921 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     1981 ttaacgcgtt gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
     2041 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
     2101 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
     2161 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
     2221 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
     2281 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     2341 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
     2401 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
     2461 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     2521 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg
     2581 ctaactagag aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag
     2641 acccaagctg gctagcgttt aaacgggccc tctagactcg agcggccgcc accatgaccc
     2701 tgagcacgga gatgtccgat gcctccggcc tcgcggagga gacagacatc gacgtggtgg
     2761 gggagggcga ggacgacgag gaggaggagg acgatgacga cgagggcggc ggcggccgcg
     2821 gcggcggcgg gtcccggttg ccgagctcgg cccagcggcg gaggcgctct tacgccgggg
     2881 aggacgatct cgaggacctg gaggaggagg acgacgatga cctgctgctg gcctcccggc
     2941 ccgccgcgtc ccccgcgcct ccgggtcctg cgcccgcccc ggggaccggg tcgggcggct
     3001 gcagcggcgc cggagcggga ggcggcgcgg gaggtggtac gggcgcgggc acgggcgggg
     3061 gcgctaagaa tccgctggtg aagccgccct actcgtacat cgcgctcatc accatggcca
     3121 tcctgcagag ccccaagaag cgcctgacgc tcagcgagat ctgcgagttc atcagcagcc
     3181 gcttccctta ctaccgggag aagttccccg cttggcagaa cagcatccgt cacaacctgt
     3241 cgctcaacga ctgcttcgtc aagatcccgc gcgaaccggg caacccgggc aagggcaact
     3301 actggacgct cgacccggag tccgcagata tgttcgacaa cggcagcttc ctgcggcgcc
     3361 gcaagcgctt caagcgccag ccgctactcg ctccccacgc ggcggccgag gcgctgctgc
     3421 tgcgcggtgc ggggccggcg gcgggcgcag gggacccggg cgccgcgctt ttcccgccgc
     3481 cgccgccacc gcccgcctgc ggctacggag cctacggctg cgcctacggc ctgcagctgc
     3541 caccctgcgc gccgccctcc gcgctcttcg ctgccgccgc cgcggcagcc gccgccgcct
     3601 tccatccgca ctcgcccccg ccgcctcctc ctcctccgcc gccgcctccc ggtgcggccg
     3661 ccgagctggc gcggaccgcc ttcggctacc ggtcacacgc gctggctgcc gccctgcccg
     3721 gccctttgca ggcggccgca gtcaaggcgg gaggcccggg cgccgcagcg ctcgcacgct
     3781 cgcccttctc catcgagagc ctcatcgggc gcactcgggg tcccgcggcc gccggggcgc
     3841 acgtctcgtc cggggccgcc agcgggaccg ccccgggacc cgggggcggg ggttgcgcag
     3901 tacaggcggc tgctggtccc gccgtcgcgc tcacccgctc gctcgtggtc gccgccgcgg
     3961 ccgccgcctc ctccgtgtcc tcgtccgccg cgctggggac tctgcaccaa gggacagcgc
     4021 tgtccagtgt ggagaacttt actgctagga tttccaattg tggatccgac tacaaagacc
     4081 atgacggtga ttataaagat catgacatcg actacaagga tgacgatgac aagtgaagct
     4141 taagtttaaa ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     4201 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     4261 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     4321 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     4381 tgggctctat gggtcgactt cgagcaactt gtttattgca gcttataatg gttacaaata
     4441 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     4501 tttgtccaaa ctcatcaatg tatcttatca tgtctggatc gtctagcatc gaagatccaa
     4561 taacttcgta tagcatacat tatacgaagt tataagtagc ttggcgtaat catggtcata
     4621 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     4681 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     4741 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     4801 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     4861 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     4921 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     4981 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     5041 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     5101 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     5161 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg
     5221 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     5281 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     5341 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     5401 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     5461 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     5521 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     5581 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5641 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5701 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     5761 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5821 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     5881 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     5941 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     6001 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     6061 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     6121 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     6181 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     6241 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     6301 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     6361 gcggcgaccg agttgctctt gcccggcgtc aacacgggat aataccgcgc cacatagcag
     6421 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6481 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6541 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6601 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6661 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6721 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     6781 cattattatc atgacattaa cctataaaaa taggcgtatc actctaggca aaatagcacc
     6841 ctcccgctcc agaacaacat acagcgcttc acagcggcag cctaacagtc agccttacca
     6901 gtaaaaaaga aaacctatta aaaaaacacc actcgacacg gcaccagctc aatcagtcac
     6961 agtgtaaaaa agggccaagt gcagagcgag tatatatagg actaaaaaat gacgtaacgg
     7021 ttaaagtcca caaaaaacac ccagaaaacc gcacgcgaac ctacgcccag aaacgaaagc
     7081 caaaaaaccc acaacttcct caaatcgtca cttccgtttt cccacgttac gtaacttccc
     7141 attttaagaa aactacaatt cccaacacat acaagttact ccgccctaaa acctacgtca
     7201 cccgccccgt tcccacgccc cgcgccacgt cacaaactcc accccctcat tatcatattg
     7261 gcttcaatcc aaaataaggt ata
//
