LOCUS       V037813                 8195 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V037813
VERSION     V037813
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     promoter        1..1179
                     /note="strong constitutive promoter for human elongation
                     factor EF-1α"
                     /label="EF-1α promoter"
     promoter        1196..1214
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     protein_bind    1293..1317
                     /bound_moiety="BP Clonase™"
                     /gene="mutant version of&nbsp;attB"
                     /note="recombination site for the Gateway® BP reaction"
                     /label="attB1"
     CDS             1348..2997
                     /EC_number=""
                     /codon_start=1
                     /gene="luc+"
                     /note="enhanced&nbsp;luc+ version of the luciferase gene"
                     /product="firefly luciferase"
                     /protein_id=""
                     /transl_table=1
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
                     /label="luciferase"
     CDS             3028..4095
                     /label="Gprc5a(NM_181444)"
                     /note="Gprc5a(NM_181444)"
                     /gene="Gprc5a"
     polyA_signal    4123..4347
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     protein_bind    4417..4450
                     /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(4908..4924)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     promoter        5398..5502
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             5503..6363
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     rep_origin      6534..7122
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     protein_bind    7410..7431
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        7446..7476
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7484..7500
                     /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"
     primer_bind     7508..7524
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    8022..8055
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (ATGTATGC) (Shaw et al., 2021)."
                     /label="loxP"
ORIGIN
        1 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
       61 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
      121 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
      181 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc
      241 gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt
      301 acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      361 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      421 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      481 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      541 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      601 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      661 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      721 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
      781 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
      841 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
      901 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
      961 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1021 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1081 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1141 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gaattagctt ggtactaata
     1201 cgactcacta tagggagacc caagctggct aggtaagctt ggtaccgagc tcggatccac
     1261 tagtccagtg tggtggaatt ctgcagatat caacaagttt gtacaaaaaa gcaggctccg
     1321 cggccgcccc cttcaccatg ggccaccatg gaagacgcca aaaacataaa gaaaggcccg
     1381 gcgccattct atccgctgga agatggaacc gctggagagc aactgcataa ggctatgaag
     1441 agatacgccc tggttcctgg aacaattgct tttacagatg cacatatcga ggtggacatc
     1501 acttacgctg agtacttcga aatgtccgtt cggttggcag aagctatgaa acgatatggg
     1561 ctgaatacaa atcacagaat cgtcgtatgc agtgaaaact ctcttcaatt ctttatgccg
     1621 gtgttgggcg cgttatttat cggagttgca gttgcgcccg cgaacgacat ttataatgaa
     1681 cgtgaattgc tcaacagtat gggcatttcg cagcctaccg tggtgttcgt ttccaaaaag
     1741 gggttgcaaa aaattttgaa cgtgcaaaaa aagctcccaa tcatccaaaa aattattatc
     1801 atggattcta aaacggatta ccagggattt cagtcgatgt acacgttcgt cacatctcat
     1861 ctacctcccg gttttaatga atacgatttt gtgccagagt ccttcgatag ggacaagaca
     1921 attgcactga tcatgaactc ctctggatct actggtctgc ctaaaggtgt cgctctgcct
     1981 catagaactg cctgcgtgag attctcgcat gccagagatc ctatttttgg caatcaaatc
     2041 attccggata ctgcgatttt aagtgttgtt ccattccatc acggttttgg aatgtttact
     2101 acactcggat atttgatatg tggatttcga gtcgtcttaa tgtatagatt tgaagaagag
     2161 ctgtttctga ggagccttca ggattacaag attcaaagtg cgctgctggt gccaacccta
     2221 ttctccttct tcgccaaaag cactctgatt gacaaatacg atttatctaa tttacacgaa
     2281 attgcttctg gtggcgctcc cctctctaag gaagtcgggg aagcggttgc caagaggttc
     2341 catctgccag gtatcaggca aggatatggg ctcactgaga ctacatcagc tattctgatt
     2401 acacccgagg gggatgataa accgggcgcg gtcggtaaag ttgttccatt ttttgaagcg
     2461 aaggttgtgg atctggatac cgggaaaacg ctgggcgtta atcaaagagg cgaactgtgt
     2521 gtgagaggtc ctatgattat gtccggttat gtaaacaatc cggaagcgac caacgccttg
     2581 attgacaagg atggatggct acattctgga gacatagctt actgggacga agacgaacac
     2641 ttcttcatcg ttgaccgcct gaagtctctg attaagtaca aaggctatca ggtggctccc
     2701 gctgaattgg aatccatctt gctccaacac cccaacatct tcgacgcagg tgtcgcaggt
     2761 cttcccgacg atgacgccgg tgaacttccc gccgccgttg ttgttttgga gcacggaaag
     2821 acgatgacgg aaaaagagat cgtggattac gtcgccagtc aagtaacaac cgcgaaaaag
     2881 ttgcgcggag gagttgtgtt tgtggacgaa gtaccgaaag gtcttaccgg aaaactcgac
     2941 gcaagaaaaa tcagagagat cctcataaag gccaagaagg gcggaaagat cgccgtgggc
     3001 ggcggaggga gtggaggagg gggaagcatg actactccta gagctgcccc tagcggttgc
     3061 cgctcagacc tggattccag gtaccacaga ctttgtgacc tggcggaagg ctggggcatc
     3121 gcgttggaaa cactggctgc cgtcggggct gtggctaccg tggcttgcat gtttgcactc
     3181 gttttcctca tctgcaaagt gcaggactcc aacaaaagaa agatgctccc cgcccagttc
     3241 ctcttcctcc tgggagtgct aggggtcttt ggcctcacct tcgccttcat catcaagctg
     3301 gatggggcca caggacccac gcgcttcttc ctcttcggag tcctcttcgc catctgcttc
     3361 tcttgcctcc tggcccacgc tttcaacttg atcaagctgg tccgagggag gaagcccctg
     3421 tcgtggcttg tgatcctcag cctggcggtg ggcttcagcc tggtccagga cgtcattgcc
     3481 attgaatacc tggtcctcac catgaacagg accaacgtca atgtcttttc tgagctgcct
     3541 gctcctcggc gcaatgagga tttcgtcatg ctcctcattt acgtgctcgt cttgatggtg
     3601 ctgaccttct tcgcatcctt cttggttttc tgtggatcct tctctggctg gaagagacac
     3661 gggttccaca tatgcttcac ctcgttcctc tccattgcca tctgggtggc ctggatcgtt
     3721 ctgctcctga ttcccgacat tgaccggaaa tgggatgata ccattctcag cacagccttg
     3781 gtggccaacg gctgggtttt cctggcgttt tatatcttgc ccgagtttcg acagctccca
     3841 aggcaacgga gccccactga ttacccagtt gaagatgctt tttgtaaacc tcagctcatg
     3901 aagcagagct atggtgtgga gaacagagcc tactcccaag aggaaatcac ccaaggtctt
     3961 gagatggggg acacactcta tgcaccttat tccacccatt ttcagctaca gaatcaccaa
     4021 aaggatttct ctatcccgag ggcccaggcc ccggccagtc cgtacaatga ctacgaaggg
     4081 cgcaaaggcg acagctaatt taaacccgct gatcagcctc gactgtgcct tctagttgcc
     4141 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     4201 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     4261 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     4321 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta
     4381 gggggtatcc ccacgcgccc tgtagcggcg cattaaataa cttcgtataa tgtatgctat
     4441 acgaagttat atcgtatgca tgctttgcat acttctgcct gctggggagc ctggggactt
     4501 tccacaccct aactgacaca cattccacag aattccttga aatacatcca caggtacacc
     4561 tccaattgac tcaaatgatg tcaattagtc tatcagaagc ttctaaagcc atgacatcat
     4621 tttctggaat tttccaagct gtttaaaggc acagtcaact tagtgtatgt aaacttctga
     4681 cccactggaa ttgtgataca gtgaattata agtgaaataa tctgtctgta aacaattgtt
     4741 ggaaaaatga cttgtgtcat gcacaaagta gatgtcctaa ctgacttgcc aaaactattg
     4801 tttgttaaca agaaatttgt ggagtagttg aaaaacgagt tttaatgact ccaacttaag
     4861 tgtatgtaaa cttccgactt caactgtagg gtaccgagct cgaattcact ggccgtcgtt
     4921 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
     4981 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     5041 ttgcgcagcc tgaatggcga atggcgcctg atgcggtatt ttctccttac gcatctgtgc
     5101 ggtatttcac accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     5161 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     5221 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     5281 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     5341 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     5401 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     5461 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     5521 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     5581 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     5641 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     5701 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     5761 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     5821 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     5881 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     5941 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     6001 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     6061 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     6121 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     6181 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     6241 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     6301 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     6361 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     6421 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     6481 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     6541 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     6601 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     6661 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     6721 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     6781 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     6841 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     6901 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     6961 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     7021 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     7081 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     7141 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     7201 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     7261 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     7321 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     7381 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     7441 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     7501 tttcacacag gaaacagcta tgaccatgat tacgccaagc ttgcatgcct gcaggtcgac
     7561 tctagaccta tacagttgaa gtcggaagtt tacatacact taagttggag tcattaaaac
     7621 tcgtttttca actactccac aaatttcttg ttaacaaaca atagttttgg caagtcagtt
     7681 aggacatcta ctttgtgcat gacacaagtc atttttccaa caattgttta cagacagatt
     7741 atttcactta taattcactg tatcacaatt ccagtgggtc agaagtttac atacactaag
     7801 ttgactgtgc ctttaaacag cttggaaaat tccagaaaat gatgtcatgg ctttagaagc
     7861 ttctgataga ctaattgaca tcatttgagt caattggagg tgtacctgtg gatgtatttc
     7921 aaggggatcc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     7981 agcggccgct agctcgagat cttaattaaa ctagtatcga tataacttcg tataatgtat
     8041 gctatacgaa gttatatgta cgggccagat atacgcgttg acattgatta ttgactaggc
     8101 ttttgcaaaa agctttgcaa agatggataa agttttaaac agagaggaat ctttgcagct
     8161 aatggacctt ctaggtcttg aaaggagtgg gaatt
//
