LOCUS       V032668                 8203 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032668
VERSION     V032668
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             931..2886
                     /label="SLC20A2(NM_001257180)"
                     /note="SLC20A2(NM_001257180)"
                     /gene="SLC20A2"
     CDS             2923..3639
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK"
                     /label="EGFP"
     CDS             3646..3735
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     polyA_signal    3770..3994
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4040..4468
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4482..4811
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4878..5672
                     /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    5846..5967
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6016..6032)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6040..6056
                     /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(6064..6094)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6109..6130
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6418..7003)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7174..8034)
                     /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(8035..8139)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cactagtcca gtgtggtgga attcgccacc atggccatgg atgagtattt gtggatggtc
      961 attttgggtt tcatcatagc tttcatcttg gccttttctg ttggtgcaaa cgatgttgcc
     1021 aactcctttg gtacagccgt gggctctggt gtggtgacct tgaggcaggc atgcatttta
     1081 gcttcaatat ttgaaaccac cggctccgtg ttactaggcg ccaaagtagg agaaaccatt
     1141 cgcaaaggta tcattgacgt gaacctgtac aacgagacgg tggagactct catggctggg
     1201 gaagttagtg ccatggttgg ttccgctgtg tggcagctga ttgcttcctt cctgaggctt
     1261 ccaatctcag gaacgcactg cattgtgggt tctactatag gattctcact ggtcgcaatc
     1321 ggtaccaaag gtgtgcagtg gatggagctt gtcaagattg ttgcttcttg gtttatatct
     1381 ccactgttgt ctggtttcat gtctggcctg ctgtttgtac tcatcagaat tttcatctta
     1441 aaaaaggaag accctgttcc caatggcctc cgggcactcc cagtattcta tgctgctacc
     1501 atagcaatca atgtcttttc catcatgtac acaggagcac cagtgctcgg ccttgttctc
     1561 cccatgtggg ccatagccct catttccttt ggtgtcgccc tcctgttcgc tttttttgtg
     1621 tggctcttcg tgtgtccgtg gatgcggagg aaaataacag gcaaattaca aaaagaaggt
     1681 gctttatcac gagtatctga cgaaagcctc agtaaggttc aggaagcaga gtccccagta
     1741 tttaaagagc taccaggtgc caaggctaat gatgacagca ccatcccgct cacgggagca
     1801 gcaggggaga cactggggac ctcggaaggc acttctgcgg gcagccaccc tcgggctgca
     1861 tacggaagag cactgtccat gacccatggc tctgtgaaat cgcccatctc caacggcacc
     1921 ttcggcttcg acggccacac caggagcgac ggtcatgtgt accacaccgt gcacaaagac
     1981 tcggggctct acaaagatct gctgcacaaa atccacatcg acaggggccc cgaggagaag
     2041 ccagcccagg aaagcaacta ccggctgctg cgccgaaaca acagttacac ctgctacacc
     2101 gcagccattt gtgggctgcc agtgcacgcc acctttcgag ctgcggactc atcggcccca
     2161 gaggacagtg agaagctggt gggcgacacc gtgtcctact ccaagaagag gctgcgctac
     2221 gacagctact cgagctactg taacgcggtg gcagaggcgg agatcgaggc ggaggagggc
     2281 ggcgtggaga tgaagctggc gtcggagctg gccgaccctg accagccgcg agaggaccct
     2341 gcagaggagg agaaggagga gaaggacgca cccgaggttc acctcctgtt ccatttcctg
     2401 caggtcctca ccgcctgttt cgggtccttt gctcacggcg gcaatgacgt gagtaatgcc
     2461 atcggtcccc tggtagcctt gtggctgatt tacaaacaag gcggggtaac gcaagaagca
     2521 gctacacccg tctggctgct gttttatgga ggagttggaa tctgcacagg cctctgggtc
     2581 tgggggagaa gagtgatcca gaccatgggg aaggacctca ctcccatcac gccgtccagc
     2641 ggcttcacga tcgagctggc ctcagccttc acagtggtga tcgcctccaa catcgggctt
     2701 ccagtcagca ccacgcactg taaggtgggc tcggtggtgg ccgtgggctg gatccgctcc
     2761 cgcaaggctg tggactggcg cctctttcgg aacatcttcg tggcctggtt cgtgaccgtc
     2821 cctgtggctg ggctgttcag cgctgctgtc atggctcttc tcatgtatgg gatccttcca
     2881 tatgtggata tcggcggtgg aggaagcggc ggaggtggat ctatggtgag caagggcgag
     2941 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     3001 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     3061 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
     3121 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
     3181 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     3241 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     3301 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     3361 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
     3421 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     3481 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     3541 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     3601 gccgccggga tcactctcgg catggacgag ctgtacaagc tcgaggaaca aaaactaata
     3661 tcagaggaag atttggaaca gaaactaatc agtgaagaag atttagagca aaagttaatt
     3721 tccgaggagg acttatgagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct
     3781 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     3841 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     3901 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     3961 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     4021 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     4081 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     4141 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     4201 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     4261 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     4321 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     4381 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     4441 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     4501 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4561 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4621 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     4681 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     4741 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     4801 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag
     4861 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     4921 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     4981 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     5041 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     5101 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     5161 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     5221 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     5281 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     5341 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     5401 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     5461 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     5521 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     5581 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     5641 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     5701 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     5761 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     5821 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     5881 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     5941 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     6001 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     6061 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     6121 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     6181 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     6241 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     6301 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     6361 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     6421 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     6481 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     6541 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     6601 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     6661 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     6721 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     6781 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     6841 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     6901 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtttttt
     6961 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     7021 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     7081 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     7141 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     7201 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     7261 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     7321 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     7381 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     7441 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     7501 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     7561 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     7621 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     7681 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     7741 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     7801 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     7861 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     7921 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     7981 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     8041 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     8101 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     8161 acctgacgtc gacggatcgg gagatctccc gatcccctat ggt
//
