LOCUS       V037221                 6958 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V037221
VERSION     V037221
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             706..1614
                     /label="Gdf15(NM_011819)"
                     /note="Gdf15(NM_011819)"
                     /gene="Gdf15"
     CDS             1621..1710
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    1734..2322
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2388..2436
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2638..3066
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3080..3409
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3476..4270
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    4446..4567
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4616..4632)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4640..4656
                     /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(4664..4694)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4709..4730
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5018..5606)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5777..6637)
                     /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"
     promoter        complement(6638..6742)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccacta gtccagtgtg gtggaattcg ccaccatggc cccgcccgcg
      721 ctccaggccc agcctccagg cggctctcaa ctgaggttcc tgctgttcct gctgctgttg
      781 ctgctgctgc tgtcatggcc atcgcagggg gacgccctgg caatgcctga acagcgaccc
      841 tccggccctg agtcccaact caacgccgac gagctacggg gtcgcttcca ggacctgctg
      901 agccggctgc atgccaacca gagccgagag gactcgaact cagaaccaag tcctgaccca
      961 gctgtccgga tactcagtcc agaggtgaga ttggggtccc acggccagct gctactccgc
     1021 gtcaaccggg cgtcgctgag tcagggtctc cccgaagcct accgcgtgca ccgagcgctg
     1081 ctcctgctga cgccgacggc ccgcccctgg gacatcacta ggcccctgaa gcgtgcgctc
     1141 agcctccggg gaccccgtgc tcccgcatta cgcctgcgcc tgacgccgcc tccggacctg
     1201 gctatgctgc cctctggcgg cacgcagctg gaactgcgct tacgggtagc cgccggcagg
     1261 gggcgccgaa gcgcgcatgc gcacccaaga gactcgtgcc cactgggtcc ggggcgctgc
     1321 tgtcacttgg agactgtgca ggcaactctt gaagacttgg gctggagcga ctgggtgctg
     1381 tccccgcgcc agctgcagct gagcatgtgc gtgggcgagt gtccccacct gtatcgctcc
     1441 gcgaacacgc atgcgcagat caaagcacgc ctgcatggcc tgcagcctga caaggtgcct
     1501 gccccgtgct gtgtcccctc cagctacacc ccggtggttc ttatgcacag gacagacagt
     1561 ggtgtgtcac tgcagactta tgatgacctg gtggcccggg gctgccactg cgctctcgag
     1621 gaacaaaaac taatatcaga ggaagatttg gaacagaaac taatcagtga agaagattta
     1681 gagcaaaagt taatttccga ggaggactta tagtgagggc ccgatatctc gacaatcaac
     1741 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     1801 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     1861 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     1921 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     1981 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     2041 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     2101 ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg
     2161 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     2221 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc
     2281 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggaaac gggggaggct
     2341 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
     2401 acagaataaa acgcacgggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg
     2461 gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt
     2521 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg
     2581 gggcggcagg ccctgccata gcagatctgc gcagctgggg ctctaggggg tatccccacg
     2641 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     2701 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     2761 tcgccggctt tccccgtcaa gctctaaatc ggggcatccc tttagggttc cgatttagtg
     2821 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     2881 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     2941 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     3001 ggattttggg gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     3061 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     3121 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     3181 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     3241 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     3301 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
     3361 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
     3421 agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat
     3481 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     3541 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     3601 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
     3661 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     3721 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     3781 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     3841 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     3901 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     3961 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
     4021 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     4081 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     4141 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     4201 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     4261 gttcttctga gcgggactct ggggttcgcg aaatgaccga ccaagcgacg cccaacctgc
     4321 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     4381 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     4441 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     4501 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     4561 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     4621 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     4681 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     4741 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     4801 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     4861 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     4921 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     4981 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     5041 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     5101 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     5161 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac
     5221 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     5281 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     5341 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     5401 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     5461 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     5521 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     5581 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     5641 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     5701 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     5761 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     5821 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     5881 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     5941 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     6001 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     6061 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     6121 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     6181 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     6241 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     6301 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     6361 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     6421 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     6481 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     6541 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     6601 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     6661 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     6721 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat
     6781 cgggagatct cccgatcccc tatggtcgac tctcagtaca atctgctctg atgccgcata
     6841 gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa
     6901 aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagg
//
