LOCUS       V028997                 6527 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028997
VERSION     V028997
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     primer_bind     141..157
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     enhancer        318..697
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        698..901
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             931..996
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             1009..1275
                     /label="Cxcl12(NM_021704)"
                     /note="Cxcl12(NM_021704)"
                     /gene="Cxcl12"
     polyA_signal    1286..1908
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     promoter        1950..2247
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             2301..3095
                     /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    3747..3881
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     promoter        complement(3887..3905)
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     primer_bind     complement(3919..3935)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    3943..3959
                     /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(3967..3997)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4012..4033
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4321..4909)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5080..5940)
                     /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(5941..6045)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      6072..6500
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attaaccatg gactacaaag accatgacgg tgattataaa
      961 gatcatgaca tcgattacaa ggatgacgat gacaagcttg cggccgcgat ggacgccaag
     1021 gtcgtcgccg tgctggccct ggtgctggcc gcgctctgca tcagtgacgg taaaccagtc
     1081 agcctgagct accgatgccc ctgccggttc ttcgagagcc acatcgccag agccaacgtc
     1141 aagcatctga aaatcctcaa cactccaaac tgtgcccttc agattgttgc acggctgaag
     1201 aacaacaaca gacaagtgtg cattgacccg aaattaaagt ggatccaaga gtacctggag
     1261 aaagctttaa acaagtaaga attccgggtg gcatccctgt gacccctccc cagtgcctct
     1321 cctggccctg gaagttgcca ctccagtgcc caccagcctt gtcctaataa aattaagttg
     1381 catcattttg tctgactagg tgtccttcta taatattatg gggtggaggg gggtggtatg
     1441 gagcaagggg caagttggga agacaacctg tagggcctgc ggggtctatt gggaaccaag
     1501 ctggagtgca gtggcacaat cttggctcac tgcaatctcc gcctcctggg ttcaagcgat
     1561 tctcctgcct cagcctcccg agttgttggg attccaggca tgcatgacca ggctcagcta
     1621 atttttgttt ttttggtaga gacggggttt caccatattg gccaggctgg tctccaactc
     1681 ctaatctcag gtgatctacc caccttggcc tcccaaattg ctgggattac aggcgtgaac
     1741 cactgctccc ttccctgtcc ttctgatttt aaaataacta taccagcagg aggacgtcca
     1801 gacacagcat aggctacctg gccatgccca accggtggga catttgagtt gcttgcttgg
     1861 cactgtcctc tcatgcgttg ggtccactca gtagatgcct gttgaattgg gtacgcggcc
     1921 agcttggctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     1981 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     2041 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     2101 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     2161 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     2221 ccagaagtag tgaggaggct tttttggagg aattgatcag cttgggatct gatcaagaga
     2281 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     2341 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     2401 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     2461 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     2521 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     2581 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     2641 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     2701 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     2761 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     2821 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     2881 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     2941 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     3001 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     3061 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     3121 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     3181 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     3241 tctcatgctg gagttcttcg cccaccccgg gctcgatccc ctcgcgagtt ggttcagctg
     3301 ctgcctgagg ctggacgacc tcgcggagtt ctaccggcag tgcaaatccg tcggcatcca
     3361 ggaaaccagc agcggctatc cgcgcatcca tgcccccgaa ctgcaggagt ggggaggcac
     3421 gatggccgct ttggtcgacc cggacgggac gctcctgcgc ctgatacaga acgaattgct
     3481 tgcaggcatc tcatgagtgt gtcttcccgt tttccgcctg aggtcactgc gtggatggag
     3541 cgctggcgcc tgctgcgcga cggcgagctg ctcaccaccc actcgccaag ctggaaccgt
     3601 aaaaaggccg cgttgctggc gtttttccat aggctccgcc gatcataatc agccatacca
     3661 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     3721 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     3781 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     3841 gtttgtccaa actcatcaat gtatcttatc atgtctggat caattcccta tagtgagtcg
     3901 tattaaattc gtaatcatgt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     3961 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     4021 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     4081 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     4141 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     4201 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     4261 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     4321 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     4381 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4441 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     4501 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     4561 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     4621 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     4681 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     4741 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     4801 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     4861 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     4921 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     4981 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     5041 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     5101 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     5161 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     5221 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     5281 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     5341 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     5401 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     5461 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     5521 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     5581 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     5641 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     5701 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     5761 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     5821 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     5881 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     5941 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6001 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     6061 agtgccacct gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     6121 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     6181 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     6241 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     6301 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     6361 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     6421 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     6481 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaattt
//
