LOCUS       V037747                 8114 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V037747
VERSION     V037747
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..4014
                     /label="Gpbar1(NM_174985)"
                     /note="Gpbar1(NM_174985)"
                     /gene="Gpbar1"
     polyA_signal    4042..4266
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     protein_bind    4336..4369
                     /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(4827..4843)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     promoter        5317..5421
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             5422..6282
                     /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      6453..7041
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     protein_bind    7329..7350
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        7365..7395
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7403..7419
                     /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     7427..7443
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7941..7974
                     /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 atgacaccca acagcactga gctgtcggcc
     3061 attcccatgg gggttctggg gctttccttg gccctggcaa gcctcatcgt catcgccaac
     3121 ctgctcctgg ccctaggcat cgccctggac cgccacttgc gcagcccacc tgctggctgc
     3181 ttcttcctaa gcctactact agccgggctg ctcacagggc tggcactgcc catgctgcct
     3241 gggctatgga gccggaacca tcagggctac tggtcctgcc tccttctcca cttgaccccc
     3301 aacttttgtt tcctttccct gcttgccaat ctgctgctgg tgcatgggga acgctacatg
     3361 gcagtgttgc agccactccg gccccatgga agtgtgcggc tagccctgtt cctcacctgg
     3421 gtcagctccc tgttctttgc cagcctgcct gctctgggct ggaaccattg gagccctgat
     3481 gccaactgca gctcccaagc tgtcttccca gccccctacc tctacctgga agtttatggc
     3541 ctcctgttgc ctgccgtggg ggccactgcc cttctctctg tccgcgtgtt ggccactgcc
     3601 caccgccagc tgtgtgagat ccgccgactg gagcgggcag tgtgccgcga tgtaccctca
     3661 accctggcta gggctctcac ctggaggcag gctagggcac aggcaggagc cacactgctc
     3721 ttcttgctgt gttgggggcc ctatgtggcc acattgctcc tgtcagtctt ggcctatgag
     3781 cgtcgcccac cactagggcc tggaactctg ttatcgctca tctcattggg cagcaccagt
     3841 gctgccgctg tgcctgtggc catggggctg ggtgatcagc gctacacagc cccctggagg
     3901 acagctgccc aaaggtgtct acgagtgctt cgaggaagag ccaagaggga caatccaggc
     3961 cccagcactg cctaccacac cagtagccaa tgcagcattg acctggactt gaattagttt
     4021 aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     4081 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     4141 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     4201 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     4261 ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct
     4321 gtagcggcgc attaaataac ttcgtataat gtatgctata cgaagttata tcgtatgcat
     4381 gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacccta actgacacac
     4441 attccacaga attccttgaa atacatccac aggtacacct ccaattgact caaatgatgt
     4501 caattagtct atcagaagct tctaaagcca tgacatcatt ttctggaatt ttccaagctg
     4561 tttaaaggca cagtcaactt agtgtatgta aacttctgac ccactggaat tgtgatacag
     4621 tgaattataa gtgaaataat ctgtctgtaa acaattgttg gaaaaatgac ttgtgtcatg
     4681 cacaaagtag atgtcctaac tgacttgcca aaactattgt ttgttaacaa gaaatttgtg
     4741 gagtagttga aaaacgagtt ttaatgactc caacttaagt gtatgtaaac ttccgacttc
     4801 aactgtaggg taccgagctc gaattcactg gccgtcgttt tacaacgtcg tgactgggaa
     4861 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     4921 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     4981 tggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg
     5041 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     5101 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     5161 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     5221 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     5281 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     5341 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     5401 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     5461 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     5521 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     5581 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     5641 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     5701 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     5761 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     5821 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     5881 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     5941 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     6001 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     6061 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     6121 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     6181 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     6241 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     6301 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     6361 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     6421 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     6481 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     6541 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     6601 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     6661 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     6721 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     6781 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     6841 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     6901 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     6961 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     7021 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     7081 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     7141 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     7201 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     7261 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     7321 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     7381 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     7441 gaccatgatt acgccaagct tgcatgcctg caggtcgact ctagacctat acagttgaag
     7501 tcggaagttt acatacactt aagttggagt cattaaaact cgtttttcaa ctactccaca
     7561 aatttcttgt taacaaacaa tagttttggc aagtcagtta ggacatctac tttgtgcatg
     7621 acacaagtca tttttccaac aattgtttac agacagatta tttcacttat aattcactgt
     7681 atcacaattc cagtgggtca gaagtttaca tacactaagt tgactgtgcc tttaaacagc
     7741 ttggaaaatt ccagaaaatg atgtcatggc tttagaagct tctgatagac taattgacat
     7801 catttgagtc aattggaggt gtacctgtgg atgtatttca aggggatcca gacatgataa
     7861 gatacattga tgagtttgga caaaccacaa ctagaatgca gcggccgcta gctcgagatc
     7921 ttaattaaac tagtatcgat ataacttcgt ataatgtatg ctatacgaag ttatatgtac
     7981 gggccagata tacgcgttga cattgattat tgactaggct tttgcaaaaa gctttgcaaa
     8041 gatggataaa gttttaaaca gagaggaatc tttgcagcta atggaccttc taggtcttga
     8101 aaggagtggg aatt
//
