LOCUS       V037477                 6637 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V037477
VERSION     V037477
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..1293
                     /label="Slc31a1(NM_175090)"
                     /note="Slc31a1(NM_175090)"
                     /gene="Slc31a1"
     CDS             1300..1389
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    1413..2001
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2067..2115
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2317..2745
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2759..3088
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3155..3949
                     /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    4125..4246
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4295..4311)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4319..4335
                     /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(4343..4373)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4388..4409
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4697..5285)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5456..6316)
                     /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(6317..6421)
                     /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 ccaccatgaa ccatatgggg
      721 atgaaccata tggagatgca ccaccatatg ggtatgaacc acacggacga caacattacc
      781 atgccacctc accaccaccc gaccacctca gcctcacact cccacggtgg aggggacagc
      841 atgatgatga tgcctatgac cttctacttt gactttaaga atgtgaatct attgttttcc
      901 ggtttggtaa tcaatacacc tggagaaatg gctggagcct tcgtggcagt gtttttacta
      961 gcaatgtttt atgaaggact caagatagcc cgagagggtc tgcttcgaaa gtctcaagtc
     1021 agcattcgct acaattccat gcctgtccca ggaccaaatg gaaccatcct aatggagaca
     1081 cacaaaactg ttgggcagca gatgctgagc ttcccccacc tcctgcagac agtgctgcac
     1141 atcatccagg tagtcatcag ctacttcctc atgctcatct tcatgaccta caatgggtac
     1201 ctatgcattg cagtagcagc aggggctggg acgggatact ttctcttcag ctggaagaag
     1261 gcagtggtgg tggacatcac agagcactgc catctcgagg aacaaaaact aatatcagag
     1321 gaagatttgg aacagaaact aatcagtgaa gaagatttag agcaaaagtt aatttccgag
     1381 gaggacttat agtgagggcc cgatatctcg acaatcaacc tctggattac aaaatttgtg
     1441 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     1501 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     1561 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     1621 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     1681 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     1741 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     1801 cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg
     1861 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     1921 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     1981 ccctttgggc cgcctccccg cctggaaacg ggggaggcta actgaaacac ggaaggagac
     2041 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacgggtg
     2101 ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc
     2161 caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc
     2221 caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag
     2281 cagatctgcg cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa
     2341 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     2401 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     2461 ctctaaatcg gggcatccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     2521 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     2581 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     2641 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgggg atttcggcct
     2701 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt
     2761 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     2821 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     2881 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     2941 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     3001 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg
     3061 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg
     3121 atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     3181 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     3241 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     3301 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg
     3361 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     3421 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     3481 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     3541 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     3601 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     3661 actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg
     3721 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     3781 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     3841 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     3901 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     3961 gggttcgcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     4021 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     4081 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     4141 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     4201 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     4261 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     4321 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     4381 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     4441 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     4501 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     4561 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     4621 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     4681 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     4741 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     4801 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     4861 ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg
     4921 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     4981 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     5041 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     5101 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     5161 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     5221 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     5281 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     5341 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     5401 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     5461 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     5521 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     5581 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     5641 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     5701 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     5761 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     5821 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     5881 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     5941 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     6001 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     6061 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     6121 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     6181 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     6241 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     6301 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     6361 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     6421 gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct
     6481 atggtcgact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc
     6541 tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca
     6601 aggcttgacc gacaattgca tgaagaatct gcttagg
//
