LOCUS       V018624                 6377 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018624
VERSION     V018624
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             1324..2247
                     /label="GDF15(NM_004864)"
                     /note="GDF15(NM_004864)"
                     /gene="GDF15"
     CDS             2248..2271
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     polyA_signal    2305..2529
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     protein_bind    2599..2632
                     /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(3090..3106)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     promoter        3580..3684
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             3685..4545
                     /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      4716..5304
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     protein_bind    5592..5613
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        5628..5658
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5666..5682
                     /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     5690..5706
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6204..6237
                     /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 gcaggctgcc
     1321 accatgcccg ggcaagaact caggacggtg aatggctctc agatgctcct ggtgttgctg
     1381 gtgctctcgt ggctgccgca tgggggcgcc ctgtctctgg ccgaggcgag ccgcgcaagt
     1441 ttcccgggac cctcagagtt gcactccgaa gactccagat tccgagagtt gcggaaacgc
     1501 tacgaggacc tgctaaccag gctgcgggcc aaccagagct gggaagattc gaacaccgac
     1561 ctcgtcccgg cccctgcagt ccggatactc acgccagaag tgcggctggg atccggcggc
     1621 cacctgcacc tgcgtatctc tcgggccgcc cttcccgagg ggctccccga ggcctcccgc
     1681 cttcaccggg ctctgttccg gctgtccccg acggcgtcaa ggtcgtggga cgtgacacga
     1741 ccgctgcggc gtcagctcag ccttgcaaga ccccaggcgc ccgcgctgca cctgcgactg
     1801 tcgccgccgc cgtcgcagtc ggaccaactg ctggcagaat cttcgtccgc acggccccag
     1861 ctggagttgc acttgcggcc gcaagccgcc agggggcgcc gcagagcgcg tgcgcgcaac
     1921 ggggaccact gtccgctcgg gcccgggcgt tgctgccgtc tgcacacggt ccgcgcgtcg
     1981 ctggaagacc tgggctgggc cgattgggtg ctgtcgccac gggaggtgca agtgaccatg
     2041 tgcatcggcg cgtgcccgag ccagttccgg gcggcaaaca tgcacgcgca gatcaagacg
     2101 agcctgcacc gcctgaagcc cgacacggtg ccagcgccct gctgcgtgcc cgccagctac
     2161 aatcccatgg tgctcattca aaagaccgac accggggtgt cgctccagac ctatgatgac
     2221 ttgttagcca aagactgcca ctgcatagat tacaaggatg acgacgataa gtgaccatgg
     2281 tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     2341 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     2401 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     2461 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     2521 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
     2581 cctgtagcgg cgcattaaat aacttcgtat aatgtatgct atacgaagtt atatcgtatg
     2641 catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc ctaactgaca
     2701 cacattccac agaattcctt gaaatacatc cacaggtaca cctccaattg actcaaatga
     2761 tgtcaattag tctatcagaa gcttctaaag ccatgacatc attttctgga attttccaag
     2821 ctgtttaaag gcacagtcaa cttagtgtat gtaaacttct gacccactgg aattgtgata
     2881 cagtgaatta taagtgaaat aatctgtctg taaacaattg ttggaaaaat gacttgtgtc
     2941 atgcacaaag tagatgtcct aactgacttg ccaaaactat tgtttgttaa caagaaattt
     3001 gtggagtagt tgaaaaacga gttttaatga ctccaactta agtgtatgta aacttccgac
     3061 ttcaactgta gggtaccgag ctcgaattca ctggccgtcg ttttacaacg tcgtgactgg
     3121 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     3181 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     3241 gaatggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     3301 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     3361 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     3421 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     3481 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     3541 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     3601 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     3661 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     3721 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     3781 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     3841 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     3901 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     3961 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     4021 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     4081 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     4141 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     4201 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     4261 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     4321 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     4381 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     4441 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     4501 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     4561 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     4621 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     4681 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     4741 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     4801 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     4861 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     4921 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     4981 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     5041 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     5101 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     5161 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     5221 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     5281 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     5341 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     5401 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     5461 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     5521 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     5581 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
     5641 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
     5701 tatgaccatg attacgccaa gcttgcatgc ctgcaggtcg actctagacc tatacagttg
     5761 aagtcggaag tttacataca cttaagttgg agtcattaaa actcgttttt caactactcc
     5821 acaaatttct tgttaacaaa caatagtttt ggcaagtcag ttaggacatc tactttgtgc
     5881 atgacacaag tcatttttcc aacaattgtt tacagacaga ttatttcact tataattcac
     5941 tgtatcacaa ttccagtggg tcagaagttt acatacacta agttgactgt gcctttaaac
     6001 agcttggaaa attccagaaa atgatgtcat ggctttagaa gcttctgata gactaattga
     6061 catcatttga gtcaattgga ggtgtacctg tggatgtatt tcaaggggat ccagacatga
     6121 taagatacat tgatgagttt ggacaaacca caactagaat gcagcggccg ctagctcgag
     6181 atcttaatta aactagtatc gatataactt cgtataatgt atgctatacg aagttatatg
     6241 tacgggccag atatacgcgt tgacattgat tattgactag gcttttgcaa aaagctttgc
     6301 aaagatggat aaagttttaa acagagagga atctttgcag ctaatggacc ttctaggtct
     6361 tgaaaggagt gggaatt
//
