LOCUS       V027281                 7460 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027281
VERSION     V027281
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             941..2950
                     /label="Slc5a2(NM_133254)"
                     /note="Slc5a2(NM_133254)"
                     /gene="Slc5a2"
     CDS             2957..3022
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3060..3284
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3330..3758
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3772..4101
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4168..4962
                     /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    5136..5257
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5306..5322)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5330..5346
                     /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(5354..5384)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5399..5420
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5708..6293)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6464..7324)
                     /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(7325..7429)
                     /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 atccgccacc atggagcaac acgtagaggc
      961 aggctctgaa cttggggagc agaaggtcct gattgataat cctgctgaca ttctggttat
     1021 cgctgcctat ttcctgctgg tcattggtgt tggcttgtgg tctatgttca gaaccaatag
     1081 aggcacagtt ggtggctact tcctggcagg acggagcatg gtgtggtggc cggttggagc
     1141 ctctctgttc gccagcaaca tcggcagcgg tcattttgtg ggcctggcag ggactggcgc
     1201 agcaagtggc ttggcggtgg ctggatttga gtggaatgcg ctcttcgtgg tgctgctcct
     1261 cggatggctt tttgtgccag tgtatctgac cgctggtgtg atcacaatgc ctcagtacct
     1321 ccgcaagcgc tttggtgggc accgtattcg cctctacctg tccgtgctct cgcttttttt
     1381 gtacattttc accaagatct cggtggatat gttctctggg gcagtattca ttcaacaggc
     1441 cctgggctgg aacatttacg cttcggtcat cgctctcttg ggcatcacca tgatttatac
     1501 tgtgacagga gggctggcgg cactgatgta cacagacact gtgcagacct tcgtcattct
     1561 tgccggggcc ttcatcctca ctggttatgc tttccatgaa gtgggcgggt actcgggtct
     1621 cttcgacaaa tacctgggag caatgacttc actgacggtg tccaaggatc catctgttgg
     1681 caacatctcc agcacctgct accagccgag gcctgactcc tatcacctgc tgcgtgaccc
     1741 tgtgacagga gacctgccat ggcctgcgct gctcctgggg cttaccattg tctcgggctg
     1801 gtattggtgc agcgatcagg taatagtgca gcggtgcctg gctggaaaga atctgactca
     1861 catcaaagct gggtgcatct tgtgtggcta cctgaagctg atgcccatgt tcctcatggt
     1921 catgccaggc atgatcagcc gcattctcta cccagatgag gtggcatgtg tggtacctga
     1981 ggtgtgtaag cgggtgtgtg gcactgaggt gggctgctct aacatcgcct acccacggct
     2041 cgtggtgaag ctcatgccca atggtctgcg cggactcatg ctggcagtca tgctggctgc
     2101 cctcatgtct tctctggcat ccatctttaa cagcagtagc acgctcttca ccatggatat
     2161 ctacacgcgc ctgcggcccc gtgcaggtga taaggagctg ctgctagttg gaaggctctg
     2221 ggtggtattc atcgtggcgg tgtccgtggc ttggctgcca gtggtgcagg cagctcaggg
     2281 tgggcagctc ttcgattaca ttcagtctgt ctccagctat ctggcacctc cagtgtctgc
     2341 ggtctttgtg cttgcactct ttgtgccccg tgttaatgag aagggagcct tctggggact
     2401 agttgggggc ctgctgatgg gcctagctcg tctcataccc gagttcttct ttggctcggg
     2461 cagctgtgtg cgaccctcag cgtgcccggc actcttctgt cgggtacact acctctattt
     2521 cgccatcatt ctcttcatct gctctggcat cctcacactt ggaatctccc tgtgcactgc
     2581 gcccatccct cagaagcatc tccatcgcct ggttttcagt ctccggcaca gcaaggagga
     2641 gcgggaggac ctggatgctg atgagttaga aggtccagcc cctgctcctg tgcagaacgg
     2701 gggccaggaa tgtgcaatgg agatggaaga ggtccagtcc ccggctccag gcctgctccg
     2761 ccggtgcctg ctttggttct gtgggatgag caagagtggg tcagggagtc ctccgcccac
     2821 taccgaggag gtggcggcaa ccaccaggcg gctggaggac atcagtgagg atccccgctg
     2881 ggcacgagta gtcaacctca atgccctact catgatgacc gtggctgtgt tcctctgggg
     2941 cttctatgca ctcgaggact acaaagacca tgacggtgat tataaagatc atgacatcga
     3001 ctacaaggat gacgatgaca agtagtgagg gcccgtttaa acccgctgat cagcctcgac
     3061 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     3121 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     3181 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     3241 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     3301 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     3361 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     3421 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     3481 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     3541 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     3601 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     3661 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     3721 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     3781 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3841 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3901 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     3961 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     4021 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     4081 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     4141 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     4201 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     4261 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     4321 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     4381 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     4441 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     4501 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     4561 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     4621 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     4681 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     4741 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     4801 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     4861 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     4921 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     4981 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     5041 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     5101 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     5161 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     5221 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     5281 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     5341 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     5401 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     5461 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     5521 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     5581 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     5641 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     5701 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     5761 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     5821 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     5881 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     5941 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     6001 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     6061 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     6121 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     6181 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     6241 gcggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6301 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6361 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6421 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6481 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6541 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6601 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6661 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6721 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     6781 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     6841 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     6901 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     6961 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     7021 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7081 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7141 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7201 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7261 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7321 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7381 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7441 gaaaagtgcc acctgacgtc
//
