LOCUS       V046434                 6536 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046434
VERSION     V046434
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             1341..1364
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             1365..2435
                     /label="G6pc1(NM_008061)"
                     /note="G6pc1(NM_008061)"
                     /gene="G6pc1"
     polyA_signal    2464..2688
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     protein_bind    2758..2791
                     /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(3249..3265)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     promoter        3739..3843
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             3844..4704
                     /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      4875..5463
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     protein_bind    5751..5772
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        5787..5817
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5825..5841
                     /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     5849..5865
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6363..6396
                     /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 gactacaagg acgacgatga caagatggag gaaggaatga
     1381 acattctcca tgactttggg atccagtcga ctcgctatct ccaagtgaat taccaagact
     1441 cccaggactg gttcatcctt gtgtctgtga ttgctgacct gaggaacgcc ttctatgtcc
     1501 tctttcccat ctggttccat cttaaagaga ctgtgggcat caatctcctc tgggtggcag
     1561 tggtcggaga ctggttcaac ctcgtcttca agtggattct gtttggacaa cgcccgtatt
     1621 ggtgggtcct ggacaccgac tactacagca acagctccgt gcctataata aagcagttcc
     1681 ctgtcacctg tgagaccgga ccaggaagtc cctctggcca tgccatgggc gcagcaggtg
     1741 tatactatgt tatggtcact tctactcttg ctatctttcg aggaaagaaa aagccaacgt
     1801 atggattccg gtgtttgaac gtcatcttgt ggttgggatt ctgggctgtg cagctgaacg
     1861 tctgtctgtc ccggatctac cttgctgctc actttcccca ccaggtcgtg gctggagtct
     1921 tgtcaggcat tgctgtggct gaaactttca gccacatccg gggcatctac aatgccagcc
     1981 tccggaagta ttgtctcatc accatcttct tgtttggttt cgcgcttgga ttctacctgc
     2041 tactaaaagg gctaggggtg gacctcctgt ggactttgga gaaagctaag agatggtgtg
     2101 agcggccaga atgggtccac cttgacacta caccctttgc cagcctcttc aaaaacctgg
     2161 gaaccctctt ggggttgggg ctggccctca actccagcat gtaccggaag agctgcaagg
     2221 gagaactcag caagttgctc ccattccgct tcgcctgcat tgtggcttcc ttggtcctcc
     2281 tgcatctctt tgactctctg aagcccccat cccaggttga gttgatcttc tacatcttgt
     2341 ctttctgcaa gagcgcaaca gttccctttg catctgtcag tcttatccca tactgcctag
     2401 cccggatcct gggacagaca cacaagaagt ctttgtaagt ttaaacccgc tgatcagcct
     2461 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     2521 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     2581 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     2641 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
     2701 aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaaata
     2761 acttcgtata atgtatgcta tacgaagtta tatcgtatgc atgctttgca tacttctgcc
     2821 tgctggggag cctggggact ttccacaccc taactgacac acattccaca gaattccttg
     2881 aaatacatcc acaggtacac ctccaattga ctcaaatgat gtcaattagt ctatcagaag
     2941 cttctaaagc catgacatca ttttctggaa ttttccaagc tgtttaaagg cacagtcaac
     3001 ttagtgtatg taaacttctg acccactgga attgtgatac agtgaattat aagtgaaata
     3061 atctgtctgt aaacaattgt tggaaaaatg acttgtgtca tgcacaaagt agatgtccta
     3121 actgacttgc caaaactatt gtttgttaac aagaaatttg tggagtagtt gaaaaacgag
     3181 ttttaatgac tccaacttaa gtgtatgtaa acttccgact tcaactgtag ggtaccgagc
     3241 tcgaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     3301 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     3361 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat
     3421 tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc
     3481 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
     3541 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     3601 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg
     3661 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     3721 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     3781 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     3841 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     3901 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     3961 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     4021 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     4081 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     4141 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     4201 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     4261 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     4321 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     4381 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     4441 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     4501 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     4561 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     4621 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     4681 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     4741 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     4801 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     4861 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     4921 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     4981 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     5041 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     5101 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     5161 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     5221 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     5281 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     5341 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     5401 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     5461 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     5521 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     5581 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     5641 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     5701 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     5761 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     5821 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     5881 cttgcatgcc tgcaggtcga ctctagacct atacagttga agtcggaagt ttacatacac
     5941 ttaagttgga gtcattaaaa ctcgtttttc aactactcca caaatttctt gttaacaaac
     6001 aatagttttg gcaagtcagt taggacatct actttgtgca tgacacaagt catttttcca
     6061 acaattgttt acagacagat tatttcactt ataattcact gtatcacaat tccagtgggt
     6121 cagaagttta catacactaa gttgactgtg cctttaaaca gcttggaaaa ttccagaaaa
     6181 tgatgtcatg gctttagaag cttctgatag actaattgac atcatttgag tcaattggag
     6241 gtgtacctgt ggatgtattt caaggggatc cagacatgat aagatacatt gatgagtttg
     6301 gacaaaccac aactagaatg cagcggccgc tagctcgaga tcttaattaa actagtatcg
     6361 atataacttc gtataatgta tgctatacga agttatatgt acgggccaga tatacgcgtt
     6421 gacattgatt attgactagg cttttgcaaa aagctttgca aagatggata aagttttaaa
     6481 cagagaggaa tctttgcagc taatggacct tctaggtctt gaaaggagtg ggaatt
//
