LOCUS       V042246                 7405 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042246
VERSION     V042246
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..2895
                     /label="Slc9a9(NM_177909)"
                     /note="Slc9a9(NM_177909)"
                     /gene="Slc9a9"
     CDS             2902..2967
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3005..3229
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3275..3703
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3717..4046
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4113..4907
                     /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    5081..5202
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5251..5267)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5275..5291
                     /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(5299..5329)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5344..5365
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5653..6238)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6409..7269)
                     /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(7270..7374)
                     /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 accatggctg ggcagcttcg gtttacgtca gggaaggatg aagatcattt tcaacaccag
     1021 ggagcagtgg agctactcgc gtttaatttt ttgctcatcc ttaccatttt aacaatctgg
     1081 ttatttaaaa atcaccgatt ccgcttcttg catgagactg gaggggcgat ggtgtatggc
     1141 ctcataatgg gactaatttt gcgatatgcc acagcgccga ctgatattga tagtggaact
     1201 gtctacaact gtgggaactt gttcttcagc ccatcaactc tactagtcaa catcaccgac
     1261 caagtttatg agtataagta ccagagagag atcaaccagc acaatatcag cccccaccaa
     1321 ggcaacgcca tactggagaa gatgacgttt gacccagaaa tcttcttcaa cgttttgctg
     1381 cctcccatca tatttcatgc aggatacagt ctgaagaaga gacacttttt tcaaaacttg
     1441 ggatccattt taacatatgc cttcttggga acagccatct cctgtgtggt catagggtta
     1501 attatgtacg gctttgtgaa agccatggtg catgctggcc agctgaagag cggagacttc
     1561 cacttcactg actgtttatt ttttggttca ctgatgtctg ctacagatcc agtgacagtg
     1621 ctggccattt tccatgaact ccacgtggac ccagacctgt acacactgct gtttggagag
     1681 agtgtactga atgatgcggt ggccatagtc ctcacatact ctatatccat ctatagtccc
     1741 aaggagaatc ccaacgcgtt tgacacggca gcgttcttcc aatccgtggg caatttccta
     1801 gggatctttg ccggctcctt tgcaatgggc tccgcatatg cagttgttac cgcactgttg
     1861 accaaattta ccaagctccg tgagttcccg atgctggaga caggcctgtt tttcctcctt
     1921 tcttggagtg ccttcctgtc tgcagaggcc gccggcttaa caggcatagt tgctgttctc
     1981 ttctgtggag tcacacaggc acattacacc tacaacaatc tgtcatcgga ttccaaactg
     2041 aggaccaaac agctgtttga atttatgaac ttcctggctg aaaacgtcat cttctgctac
     2101 atgggcctgg ctctgttcac cttccagaat cacattttca atgctctttt tatactcgga
     2161 gcctttctag caatttttgt tgccagagcc tgcaacattt accccctctc cttcctcctg
     2221 aatctgggca gaaagcagaa gattccctgg aacttccagc acatgatgat gttttcaggt
     2281 ttgcgaggtg ccattgcctt cgccctggcc atccgcaaca cggaatctca gcccaaacag
     2341 atgatgttca ccaccacact gctccttgtg ttcttcactg tctgggtgtt tggaggagga
     2401 acgaccccca tgctgacgtg gcttcagatc agagttggtg tggacctgga tgaaagtctg
     2461 aaggaagagc cctcctcaca gcaggaagca aataagttgg acaaaaacat gacgaagaca
     2521 gagagtgcac agctcttcag aatgtggtat ggatttgacc acaaatacct gaaacccatc
     2581 ctaacacact caggccctcc actgaccacc accttgccag cgtggtgtgg cccagtctcc
     2641 aggctgctca ccagtccgca ggcctatggg gaacaactga aagaggatga cgtggaatgt
     2701 atcgtcaacc aagatgaact ggccatgaat taccaggagc agagcccttc acccagcagt
     2761 cccaccacca agctagctct ggaccagaag tcttcaggcc agactccagg caaggagaat
     2821 atctacgagg gagatctcgg cctgggaggc tatgatctca aactggagca gactcggggt
     2881 caaccccaga tggacctcga ggactacaaa gaccatgacg gtgattataa agatcatgac
     2941 atcgactaca aggatgacga tgacaagtag tgagggcccg tttaaacccg ctgatcagcc
     3001 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     3061 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     3121 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     3181 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
     3241 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     3301 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     3361 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     3421 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     3481 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     3541 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     3601 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     3661 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
     3721 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3781 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     3841 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     3901 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     3961 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     4021 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
     4081 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     4141 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     4201 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     4261 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     4321 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     4381 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     4441 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     4501 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     4561 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     4621 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     4681 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     4741 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     4801 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     4861 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     4921 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     4981 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     5041 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     5101 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     5161 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     5221 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     5281 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     5341 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     5401 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     5461 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     5521 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     5581 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     5641 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     5701 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     5761 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     5821 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     5881 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     5941 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     6001 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     6061 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     6121 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     6181 tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     6241 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     6301 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     6361 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     6421 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     6481 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     6541 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     6601 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     6661 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     6721 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     6781 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     6841 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     6901 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     6961 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     7021 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     7081 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     7141 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     7201 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     7261 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     7321 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     7381 tccccgaaaa gtgccacctg acgtc
//
