LOCUS       V039468                 7183 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V039468
VERSION     V039468
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             964..2673
                     /label="SLC47A1(NM_018242)"
                     /note="SLC47A1(NM_018242)"
                     /gene="SLC47A1"
     CDS             2680..2745
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2783..3007
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3053..3481
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3495..3824
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3891..4685
                     /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    4859..4980
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5029..5045)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5053..5069
                     /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(5077..5107)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5122..5143
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5431..6016)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6187..7047)
                     /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(7048..7152)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc
      961 accatggaag ctcctgagga gcccgcgcca gtgcgcggag gcccggaggc cacccttgag
     1021 gtccgtgggt cgcgctgctt gcggctgtcc gccttccgag aagagctgcg ggcgctcttg
     1081 gtcctggctg gccccgcgtt cttggttcag ctgatggtgt tcctgatcag cttcataagc
     1141 tccgtgttct gtggccacct gggcaagctg gagctggatg cagtcacgct ggcaatcgcg
     1201 gttatcaatg tcactggtgt ctcagtggga ttcggcttat cttctgcctg tgacaccctc
     1261 atctcccaga cgtacgggag ccagaacctg aagcacgtgg gcgtgatcct gcagcggagt
     1321 gcgctcgtcc tgctcctctg ctgcttcccc tgctgggcgc tctttctcaa cacccagcac
     1381 atcctgctgc tcttcaggca ggacccagat gtgtccaggc ttacccagac ctatgtcacg
     1441 atcttcattc cagctcttcc tgcaaccttt ctttatatgt tacaagttaa atatttgctc
     1501 aaccagggaa ttgtactgcc ccagatcgta actggagttg cagccaacct tgtcaatgcc
     1561 ctcgccaact atctgtttct ccatcaactg catcttgggg tgataggctc tgcactggca
     1621 aacttgattt cccagtacac cctggctcta ctcctctttc tctacatcct cgggaaaaaa
     1681 ctgcatcaag ctacatgggg aggctggtcc ctcgagtgcc tgcaggactg ggcctccttc
     1741 ctccgcctgg ccatccccag catgctcatg ctgtgcatgg agtggtgggc ctatgaggtc
     1801 gggagcttcc tcagtggcat cctcggcatg gtggagctgg gcgctcagtc catcgtgtat
     1861 gaactggcca tcattgtgta catggtccct gcaggcttca gtgtggctgc cagtgtccgg
     1921 gtaggaaacg ctctgggtgc tggagacatg gagcaggcac ggaagtcctc taccgtttcc
     1981 ctgctgatta cagtgctctt tgctgtagcc ttcagtgtcc tgctgttaag ctgtaaggat
     2041 cacgtggggt acatttttac taccgaccga gacatcatta atctggtggc tcaggtggtt
     2101 ccaatttatg ctgtttccca cctctttgaa gctcttgctt gcacgagtgg tggtgttctg
     2161 agggggagtg gaaatcagaa ggttggagcc attgtgaata ccattgggta ctatgtggtt
     2221 ggcctcccca tcgggatcgc gctgatgttt gcaaccacac ttggagtgat gggtctgtgg
     2281 tcagggatca tcatctgtac agtctttcaa gctgtgtgtt ttctaggctt tattattcag
     2341 ctaaattgga aaaaagcctg tcagcaggct caggtacacg ccaatttgaa agtaaacaac
     2401 gtgcctcgga gtgggaattc tgctctccct caggatccgc ttcacccagg gtgccctgaa
     2461 aaccttgaag gaattttaac gaacgatgtt ggaaagacag gcgagcctca gtcagatcag
     2521 cagatgcgcc aagaagaacc tttgccggaa catccacagg acggcgctaa attgtccagg
     2581 aaacagctgg tgctgcggcg agggcttctg ctcctggggg tcttcttaat cttgctggtg
     2641 gggattttag tgagattcta tgtcagaatt cagctcgagg actacaaaga ccatgacggt
     2701 gattataaag atcatgacat cgactacaag gatgacgatg acaagtagtg agggcccgtt
     2761 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     2821 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     2881 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     2941 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     3001 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     3061 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     3121 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     3181 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     3241 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     3301 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     3361 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     3421 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     3481 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     3541 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     3601 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     3661 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     3721 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     3781 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     3841 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
     3901 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     3961 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     4021 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     4081 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     4141 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     4201 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     4261 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     4321 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     4381 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     4441 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     4501 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     4561 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     4621 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     4681 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     4741 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     4801 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     4861 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     4921 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     4981 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     5041 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     5101 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     5161 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     5221 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     5281 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5341 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5401 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5461 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5521 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5581 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     5641 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5701 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5761 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5821 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     5881 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5941 cggcaaacaa accaccgctg gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag
     6001 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6061 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6121 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     6181 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     6241 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     6301 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     6361 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     6421 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     6481 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     6541 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     6601 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     6661 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     6721 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     6781 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     6841 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     6901 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     6961 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     7021 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     7081 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     7141 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc
//
