LOCUS       V023008                 7798 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023008
VERSION     V023008
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             649..1365
                     /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             1399..2475
                     /label="Cxcr4(NM_009911)"
                     /note="Cxcr4(NM_009911)"
                     /gene="Cxcr4"
     CDS             2482..2547
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2574..3162
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3228..3276
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3478..3906
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3920..4249
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4316..5110
                     /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    5286..5407
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5456..5472)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5480..5496
                     /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(5504..5534)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5549..5570
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5858..6446)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6617..7477)
                     /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(7478..7582)
                     /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 gcgccaccat ggtgagcaag
      661 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
      721 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
      781 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
      841 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
      901 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
      961 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     1021 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     1081 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
     1141 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     1201 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     1261 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     1321 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagggagg agggggaagc
     1381 ggtaccgagc tcggatccat ggaaccgatc agtgtgagta tatacacttc tgataactac
     1441 tctgaagaag tgggttctgg agactatgac tccaacaagg aaccctgctt ccgggatgaa
     1501 aacgtccatt tcaataggat cttcctgccc accatctact tcatcatctt cttgactggc
     1561 atagtcggca atggattggt gatcctggtc atgggttacc agaagaagct aaggagcatg
     1621 acggacaagt accggctgca cctgtcagtg gctgacctcc tctttgtcat cacactcccc
     1681 ttctgggcag ttgatgccat ggctgactgg tactttggga aatttttgtg taaggctgtc
     1741 catatcatct acactgtcaa cctctacagc agcgttctca tcctggcctt catcagcctg
     1801 gaccggtacc tcgctattgt ccacgccacc aacagtcaga ggccaaggaa actgctggct
     1861 gaaaaggcag tctatgtggg cgtctggatc ccagccctcc tcctgactat acctgacttc
     1921 atctttgccg acgtcagcca gggggacatc agtcaggggg atgacaggta catctgtgac
     1981 cgcctttacc ccgatagcct gtggatggtg gtgtttcaat tccagcatat aatggtgggt
     2041 ctcgtcctgc ccggcatcgt catcctctcc tgttactgca tcatcatctc taagctgtca
     2101 cactccaagg gccaccagaa gcgcaaggcc ctcaagacga cagtcatcct catcctagct
     2161 ttctttgcct gctggctgcc atattatgtg gggatcagca tcgactcctt catccttttg
     2221 ggggtcatca agcaaggatg tgacttcgag agcatcgtgc acaagtggat ctccatcaca
     2281 gaggccctcg ccttcttcca ctgttgcctg aaccccatcc tctatgcctt cctcggggcc
     2341 aagttcaaaa gctctgccca gcatgcactc aactccatga gcagaggctc cagcctcaag
     2401 atcctttcca aaggaaagcg gggtggacac tcttccgtct ccacggagtc agaatcctcc
     2461 agttttcact ccagctcgag agactacaaa gaccatgacg gtgattataa agatcatgac
     2521 atcgactaca aggatgacga tgacaagtag tctagagggc ccgatatctc gacaatcaac
     2581 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     2641 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     2701 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     2761 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     2821 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     2881 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     2941 ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg
     3001 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     3061 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc
     3121 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggaaac gggggaggct
     3181 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
     3241 acagaataaa acgcacgggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg
     3301 gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt
     3361 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg
     3421 gggcggcagg ccctgccata gcagatctgc gcagctgggg ctctaggggg tatccccacg
     3481 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     3541 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     3601 tcgccggctt tccccgtcaa gctctaaatc ggggcatccc tttagggttc cgatttagtg
     3661 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     3721 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     3781 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     3841 ggattttggg gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     3901 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     3961 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     4021 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     4081 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     4141 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
     4201 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
     4261 agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat
     4321 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     4381 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     4441 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
     4501 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     4561 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     4621 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     4681 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     4741 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     4801 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
     4861 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     4921 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     4981 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     5041 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     5101 gttcttctga gcgggactct ggggttcgcg aaatgaccga ccaagcgacg cccaacctgc
     5161 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     5221 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     5281 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     5341 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     5401 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     5461 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     5521 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     5581 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     5641 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     5701 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     5761 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     5821 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     5881 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     5941 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     6001 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac
     6061 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     6121 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     6181 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     6241 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     6301 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     6361 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     6421 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     6481 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     6541 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     6601 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     6661 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     6721 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     6781 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     6841 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     6901 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     6961 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     7021 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     7081 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     7141 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     7201 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     7261 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     7321 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     7381 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     7441 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     7501 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     7561 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat
     7621 cgggagatct cccgatcccc tatggtcgac tctcagtaca atctgctctg atgccgcata
     7681 gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa
     7741 aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagg
//
