LOCUS       V031862                 7292 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031862
VERSION     V031862
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             2712..4070
                     /label="Acot2(NM_138907)"
                     /note="Acot2(NM_138907)"
                     /gene="Acot2"
     CDS             4077..4142
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    4177..4401
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     polyA_signal    4415..4549
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     protein_bind    complement(4569..4602)
                     /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(4624..4640)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4648..4664
                     /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(4672..4702)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4717..4738
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5026..5614)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5785..6645)
                     /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(6646..6750)
                     /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 actgtgctgg
     2701 atatcgccac catggtggcc tcgtctttcg ctgtcctgag agcaagcagg ttgtgccaat
     2761 ggggttggaa gagctggacg cagctgtcag gtcctccgcc gctcagcacc ggtggccgga
     2821 ccacttttgc gcggacaaat gctacgctga gcctggagcc cgggagccgc agctgctggg
     2881 acgagccgtt gagcatcacc gtgcgcggcc tggcccccga gcagcccgtc acgctgcgcg
     2941 cggccctgcg tgacgagaag ggcgcgctct tccgagccca cgcgcgctac cgcgccgacg
     3001 ccggtggtga gctggacctg gcgcgcgctc ccgcgctggg cggcagcttc acggggctcg
     3061 agcccatggg gctgatctgg gccatggagc ccgaacggcc tctctggcgc ctggtcaagc
     3121 gcgacgtgca gaagccttat gtggtggagc tggaggtgct ggacggacac gagcccgacg
     3181 gcggtcagcg gctggcacag gcagtgcacg agcgtcactt catggctcca ggggtgcggc
     3241 gcgtgcccgt gcgcgacggc cgggtgcgcg ccacgctctt cctgccccca gaacctgggc
     3301 cctttcctgg aatcatagac ctttttggag ttggaggcgg ccttctggag taccgggcga
     3361 gtctgctggc tgggaagggt tttgccgtca tggctctggc ttattacaac tacgacgacc
     3421 tccccaagac catggaaacc atgcgcattg agtactttga agaagccgtg aactacctgc
     3481 gtggccaccc tgaggtaaaa ggaccaggaa ttgggctgct tgggatttcc aaagggggtg
     3541 aacttggcct tgctatggcc tccttcctga agggcatcac ggctgctgtt gtcatcaatg
     3601 gctccgtggc tgctgttggg aacaccgtat gctacaagga tgagactata ccccctgtgt
     3661 cccttctgag agacaaagtc aaaatgacca aagatggtct cttggatgtc gtggaagctc
     3721 tgcaaagccc tttggtagac aagaagagct tcatccctgt ggaaaggtct gacacgacct
     3781 tcctgttcct cgttggtcag gatgaccaca actggaagag cgagttctat gccagagagg
     3841 cctccaaacg cttgcaggcc cacgggaaag agaagcccca gatcatctgc tacccagaag
     3901 cagggcacta tatcgagcct ccttacttcc cactgtgcag cgctggcatg cacctcctgg
     3961 tgggtgctaa catcaccttt ggaggggagc ctaagcctca ctctgtggcc cagttggatg
     4021 catggcagca actccagact ttcttccaca aacagttgag tggtaagagt ggatccgact
     4081 acaaagacca tgacggtgat tataaagatc atgacatcga ctacaaggat gacgatgaca
     4141 agtgaagctt aagtttaaac cgctgatcag cctcgactgt gccttctagt tgccagccat
     4201 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     4261 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     4321 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     4381 gggatgcggt gggctctatg ggtcgacttc gagcaacttg tttattgcag cttataatgg
     4441 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     4501 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatcg tctagcatcg
     4561 aagatccaat aacttcgtat agcatacatt atacgaagtt ataagtagct tggcgtaatc
     4621 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     4681 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     4741 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     4801 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     4861 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     4921 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     4981 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5041 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5101 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5161 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5221 atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5281 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5341 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5401 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     5461 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     5521 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     5581 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     5641 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     5701 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     5761 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     5821 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     5881 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     5941 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     6001 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     6061 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     6121 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     6181 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     6241 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     6301 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     6361 atagtgtatg cggcgaccga gttgctcttg cccggcgtca acacgggata ataccgcgcc
     6421 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     6481 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     6541 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     6601 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     6661 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     6721 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     6781 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca ctctaggcaa
     6841 aatagcaccc tcccgctcca gaacaacata cagcgcttca cagcggcagc ctaacagtca
     6901 gccttaccag taaaaaagaa aacctattaa aaaaacacca ctcgacacgg caccagctca
     6961 atcagtcaca gtgtaaaaaa gggccaagtg cagagcgagt atatatagga ctaaaaaatg
     7021 acgtaacggt taaagtccac aaaaaacacc cagaaaaccg cacgcgaacc tacgcccaga
     7081 aacgaaagcc aaaaaaccca caacttcctc aaatcgtcac ttccgttttc ccacgttacg
     7141 taacttccca ttttaagaaa actacaattc ccaacacata caagttactc cgccctaaaa
     7201 cctacgtcac ccgccccgtt cccacgcccc gcgccacgtc acaaactcca ccccctcatt
     7261 atcatattgg cttcaatcca aaataaggta ta
//
