LOCUS       V045545                 7560 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045545
VERSION     V045545
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..2240
                     /label="Slc17a5(NM_172773)"
                     /note="Slc17a5(NM_172773)"
                     /gene="Slc17a5"
     CDS             2247..2312
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2336..2924
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2990..3038
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3240..3668
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3682..4011
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4078..4872
                     /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    5048..5169
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5218..5234)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5242..5258
                     /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(5266..5296)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5311..5332
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5620..6208)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6379..7239)
                     /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(7240..7344)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatgag gcccctgctt cggggtccgg
      781 cgggaaacga cgatgaggag agctcggaca gcaccccgct cctgccgggc gcccggcaga
      841 ccgaagcggc tccagtgtgc tgctctgctc ggtacaactt agcgattttg gcgttctgtg
      901 gtttcttcgt tctctatgcc ttacgggtga acctgagtgt tgcgttagtg gacatggtag
      961 attcaaatac aactctgact gataatagaa cgtctaagga gtgtgcggaa cattctgccc
     1021 ccataaaagt tcaccacaat cacacaggta aaaagtacaa gtgggatgca gaaactcaag
     1081 ggtggattct cggctctttt ttttacggct acatcgtcac ccagattccc ggtgggtaca
     1141 ttgccagcag ggtcggaggg aagctgctgc tgggcctggg catcttaggc acctccgtct
     1201 tcaccctgtt cacaccgctg gccgcagact taggcgtggt gactctcgtt gtgcttagag
     1261 cgctggaagg actgggagag ggtgttacgt ttccagctat gcacgccatg tggtcttcct
     1321 gggctccccc tctggaaaga agcaagcttc ttaccatttc ctatgcggga gcacagcttg
     1381 ggacagtgat ctcacttcct ctttccggaa taatatgcta ctatatgaac tggacttacg
     1441 tcttctatct ttttggtata gttggaattg tctggtttat tttatggatg tggatagtca
     1501 gtgatacacc agaaactcac aagacaatct cccattatga aaaagaatac attgtttcat
     1561 cattaaaaaa tcagctttct tcgcagaagg tggtgccgtg ggggtccatc ttgaagtcac
     1621 tgccactttg ggcaattgtg gtagcacatt tctcctacaa ctggtctttt tacaccttat
     1681 tgacgctact gccaacttat atgaaggaaa tcctaaggtt caatgttcaa gagaacgggt
     1741 ttttatctgc attgccttat tttggctgtt ggttatgcat gatcctctgt ggtcaagccg
     1801 ctgactattt aagggtcaag tggaactttt caactataag cgttcggaga atttttagcc
     1861 tcgtaggaat ggttggccct gcggttttcc tagtcgcggc tggatttata ggctgtgact
     1921 attccttggc cgttgcgttc ctaaccatat ccacgacgct gggaggcttc gcctcttctg
     1981 gatttagcat caaccatctg gatatcgctc cttcgtatgc tggcatcctc ttgggcatca
     2041 caaacacgtt tgccactatt cccgggatga cgggacccat cattgctaaa agtctgaccc
     2101 ctgataacac tattagggaa tggcagactg tcttctgtat cgctgctgct attaacgtgt
     2161 ttggggccat tttcttcacg ctgtttgcca aaggggaagt gcagagctgg gctctcagtg
     2221 accaccacgg acacagaaac ggatccgact acaaagacca tgacggtgat tataaagatc
     2281 atgacatcga ctacaaggat gacgatgaca agtagtgagg gcccgatatc tcgacaatca
     2341 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     2401 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     2461 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     2521 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     2581 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     2641 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     2701 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg
     2761 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     2821 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     2881 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa acgggggagg
     2941 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     3001 agacagaata aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     3061 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     3121 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     3181 cggggcggca ggccctgcca tagcagatct gcgcagctgg ggctctaggg ggtatcccca
     3241 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     3301 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     3361 gttcgccggc tttccccgtc aagctctaaa tcggggcatc cctttagggt tccgatttag
     3421 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     3481 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     3541 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     3601 agggattttg gggatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     3661 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3721 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3781 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3841 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3901 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     3961 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     4021 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     4081 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     4141 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     4201 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     4261 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     4321 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     4381 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     4441 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     4501 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     4561 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     4621 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     4681 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     4741 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     4801 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     4861 gagttcttct gagcgggact ctggggttcg cgaaatgacc gaccaagcga cgcccaacct
     4921 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     4981 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     5041 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     5101 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     5161 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     5221 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     5281 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     5341 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     5401 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     5461 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     5521 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     5581 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5641 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5701 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5761 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc
     5821 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5881 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5941 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     6001 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     6061 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     6121 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     6181 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     6241 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     6301 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     6361 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     6421 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     6481 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     6541 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     6601 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     6661 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     6721 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     6781 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     6841 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     6901 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     6961 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     7021 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     7081 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     7141 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     7201 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     7261 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     7321 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
     7381 atcgggagat ctcccgatcc cctatggtcg actctcagta caatctgctc tgatgccgca
     7441 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc
     7501 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
//
