LOCUS       V019913                 8912 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019913
VERSION     V019913
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             683..3592
                     /label="SLC26A8(NM_052961)"
                     /note="SLC26A8(NM_052961)"
                     /gene="SLC26A8"
     CDS             3599..3664
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3688..4276
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4342..4390
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4592..5020
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5034..5363
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5430..6224
                     /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    6400..6521
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6570..6586)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6594..6610
                     /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(6618..6648)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6663..6684
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6972..7560)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7731..8591)
                     /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(8592..8696)
                     /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 ggatccgcca ccatggcaca actagagagg agcgccatct ctggcttcag
      721 ctctaagtcc aggcgaaact cattcgcata tgatgttaag cgtgaagtat acaatgagga
      781 gacctttcaa caggaacaca aaaggaaggc ctcctcttct gggaacatga acatcaacat
      841 caccaccttc agacaccacg tccagtgccg ctgctcatgg cacaggttcc tacgatgcgt
      901 gcttacaatc tttcccttcc tagaatggat gtgtatgtat cgattaaagg attggcttct
      961 gggagactta cttgctggta taagtgttgg ccttgtgcaa gttccccaag gcctgacact
     1021 tagtttgctg gcaaggcaac tgattcctcc tctcaacatc gcttatgcag ctttctgttc
     1081 ttcggtaatc tatgtaattt ttggatcgtg tcatcaaatg tccattggtt ccttcttcct
     1141 ggtgagtgct ctgctgatca acgttctgaa agtgagccca ttcaacaacg gtcaactggt
     1201 catgggatct ttcgtcaaga atgagttttc ggccccctcc taccttatgg gctataataa
     1261 atccttgagt gtggtggcaa ccacaacttt tctgactggg attattcagc taataatggg
     1321 cgtattgggt ttgggcttca ttgccactta ccttccggag tctgcaatga gtgcttacct
     1381 ggctgctgtg gcacttcata tcatgctgtc ccagctgact ttcatctttg ggattatgat
     1441 tagtttccat gccggtccca tctccttctt ctatgacata attaattact gtgtagctct
     1501 cccaaaagcg aattccacca gcattctagt atttctaact gttgttgttg ctctgcgaat
     1561 caacaaatgt atcagaattt ctttcaatca gtatcccatt gagtttccca tggaattatt
     1621 tctgattatt ggcttcactg tgattgcaaa caagataagc atggccacag aaaccagcca
     1681 gacgcttatt gacatgattc cttatagctt tctgcttcct gtaacaccag atttcagcct
     1741 tcttcccaag ataattttac aagccttctc cttatctttg gtgagctcct ttctgctcat
     1801 atttctgggc aagaagattg ccagtcttca caattacagt gtcaattcca accaggattt
     1861 aatagccatc ggcctttgca atgtcgtcag ttcatttttc agatcttgtg tgtttactgg
     1921 tgctattgct aggactatta tccaggataa atctggagga agacaacagt ttgcatctct
     1981 ggtaggcgca ggtgtgatgc tgctcctgat ggtgaagatg ggacactttt tctacacact
     2041 gccaaatgct gtgctggctg gtattattct gagcaacgtc attccctacc ttgaaaccat
     2101 ttctaaccta cccagcctgt ggaggcagga ccaatatgac tgtgctcttt ggatgatgac
     2161 attctcatct tcaattttcc tgggactgga cattggacta attatctcag tagtttctgc
     2221 tttcttcatc accactgttc gttcacacag agctaagatt cttctcctgg gtcaaatccc
     2281 taacaccaac atttatagaa gcatcaatga ttatcgggag atcatcacca ttcctggggt
     2341 gaaaatcttc cagtgctgca gctcaattac atttgtaaat gtttactacc taaagcataa
     2401 gctgttaaaa gaggttgata tggtaaaggt gcctcttaaa gaagaagaaa ttttcagctt
     2461 gtttaattca agtgacacca atctacaagg aggaaagatt tgcaggtgtt tctgcaactg
     2521 tgatgatctg gagccgctgc ccaggattct ttacacagag cgatttgaaa ataaactgga
     2581 tcccgaagca tcctccatta acctgattca ctgctcacat tttgagagca tgaacacaag
     2641 ccaaactgca tccgaagacc aagtgccata cacagtatcg tccgtgtctc agaaaaatca
     2701 agggcaacag tatgaggagg tggaggaagt ttggcttcct aataactcat caagaaacag
     2761 ctcaccagga ctgcctgatg tggcggaaag ccaggggagg agatcactca tcccttactc
     2821 agatgcgtct ctactgccca gtgtccacac catcatcctg gatttctcca tggtacacta
     2881 cgtggattca cgggggttag tcgtattaag acagatatgc aatgcctttc aaaacgccaa
     2941 cattttgata ctcattgcag ggtgtcactc ttccatagtc agggcatttg agaggaatga
     3001 tttctttgac gctggcatca ccaagaccca gctgttcctc agcgttcacg acgccgtgct
     3061 gtttgccttg tcaaggaagg tcataggctc ctctgagtta agcatcgatg aatccgagac
     3121 agtgatacgg gaaacctact cagaaacaga caagaatgac aattcaagat ataaaatgag
     3181 cagcagtttt ctaggaagcc aaaaaaatgt aagtccaggc ttcatcaaga tccaacagcc
     3241 tgtagaagag gagtcggagt tggatttgga gctggaatca gaacaagagg ctgggctggg
     3301 tctggaccta gacctggatc gggagctgga gcctgaaatg gagcccaagg ctgagaccga
     3361 gaccaagacc cagaccgaga tggagcccca gcctgagact gagcctgaga tggagcccaa
     3421 ccccaaatct aggccaagag ctcacacttt tcctcagcag cgttactggc ctatgtatca
     3481 tccgtctatg gcttccaccc agtctcagac tcagactcgg acatggtcag tggagaggag
     3541 acgccatcct atggattcat actcaccaga gggcaacagc aatgaagatg tcctcgagga
     3601 ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg atgacgatga
     3661 caagtagtga gggcccgata tctcgacaat caacctctgg attacaaaat ttgtgaaaga
     3721 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     3781 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     3841 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     3901 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     3961 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     4021 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     4081 aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
     4141 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     4201 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     4261 tgggccgcct ccccgcctgg aaacggggga ggctaactga aacacggaag gagacaatac
     4321 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac gggtgttggg
     4381 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     4441 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     4501 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcagat
     4561 ctgcgcagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     4621 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     4681 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     4741 aatcggggca tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     4801 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     4861 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     4921 aaccctatct cggtctattc ttttgattta taagggattt tggggatttc ggcctattgg
     4981 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     5041 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     5101 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     5161 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     5221 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     5281 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     5341 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     5401 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     5461 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     5521 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     5581 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     5641 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     5701 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     5761 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     5821 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     5881 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     5941 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     6001 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     6061 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     6121 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     6181 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     6241 cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     6301 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     6361 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     6421 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     6481 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     6541 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     6601 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     6661 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     6721 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     6781 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     6841 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     6901 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     6961 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     7021 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     7081 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     7141 ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag
     7201 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     7261 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     7321 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     7381 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     7441 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     7501 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     7561 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     7621 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     7681 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     7741 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     7801 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     7861 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     7921 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     7981 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     8041 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     8101 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     8161 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     8221 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     8281 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     8341 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     8401 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     8461 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     8521 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     8581 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     8641 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     8701 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
     8761 cgactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
     8821 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
     8881 tgaccgacaa ttgcatgaag aatctgctta gg
//
