LOCUS       V047175                 6634 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V047175
VERSION     V047175
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..2124
                     /label="CX3CR1(NM_001171174)"
                     /note="CX3CR1(NM_001171174)"
                     /gene="CX3CR1"
     CDS             2131..2196
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2234..2458
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2504..2932
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2946..3275
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3342..4136
                     /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    4310..4431
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4480..4496)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4504..4520
                     /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(4528..4558)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4573..4594
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4882..5467)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5638..6498)
                     /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(6499..6603)
                     /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 accatgagag aacccctgga ggcgtttaag ttggcagact tggatttcag gaagagctct
     1021 ctggcttctg ggtggagaat ggccagtggg gccttcacca tggatcagtt ccctgaatca
     1081 gtgacagaaa actttgagta cgatgatttg gctgaggcct gttatattgg ggacatcgtg
     1141 gtctttggga ctgtgttcct gtccatattc tactccgtca tctttgccat tggcctggtg
     1201 ggaaatttgt tggtagtgtt tgccctcacc aacagcaaga agcccaagag tgtcaccgac
     1261 atttacctcc tgaacctggc cttgtctgat ctgctgtttg tagccacttt gcccttctgg
     1321 actcactatt tgataaatga aaagggcctc cacaatgcca tgtgcaaatt cactaccgcc
     1381 ttcttcttca tcggcttttt tggaagcata ttcttcatca ccgtcatcag cattgatagg
     1441 tacctggcca tcgtcctggc cgccaactcc atgaacaacc ggaccgtgca gcatggcgtc
     1501 accatcagcc taggcgtctg ggcagcagcc attttggtgg cagcacccca gttcatgttc
     1561 acaaagcaga aagaaaatga atgccttggt gactaccccg aggtcctcca ggaaatctgg
     1621 cccgtgctcc gcaatgtgga aacaaatttt cttggcttcc tactccccct gctcattatg
     1681 agttattgct acttcagaat catccagacg ctgttttcct gcaagaacca caagaaagcc
     1741 aaagccatta aactgatcct tctggtggtc atcgtgtttt tcctcttctg gacaccctac
     1801 aacgttatga ttttcctgga gacgcttaag ctctatgact tctttcccag ttgtgacatg
     1861 aggaaggatc tgaggctggc cctcagtgtg actgagacgg ttgcatttag ccattgttgc
     1921 ctgaatcctc tcatctatgc atttgctggg gagaagttca gaagatacct ttaccacctg
     1981 tatgggaaat gcctggctgt cctgtgtggg cgctcagtcc acgttgattt ctcctcatct
     2041 gaatcacaaa ggagcaggca tggaagtgtt ctgagcagca attttactta ccacacgagt
     2101 gatggagatg cattgctcct tctcctcgag gactacaaag accatgacgg tgattataaa
     2161 gatcatgaca tcgactacaa ggatgacgat gacaagtagt gagggcccgt ttaaacccgc
     2221 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     2281 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     2341 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     2401 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     2461 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     2521 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     2581 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     2641 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     2701 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     2761 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     2821 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     2881 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     2941 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3001 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     3061 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     3121 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     3181 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     3241 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     3301 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     3361 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     3421 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     3481 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     3541 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     3601 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     3661 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     3721 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     3781 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     3841 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     3901 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     3961 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     4021 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     4081 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     4141 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     4201 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     4261 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     4321 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     4381 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     4441 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     4501 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     4561 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     4621 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     4681 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     4741 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     4801 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     4861 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     4921 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     4981 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     5041 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     5101 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     5161 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     5221 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     5281 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     5341 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     5401 aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     5461 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     5521 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     5581 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     5641 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     5701 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     5761 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     5821 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     5881 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     5941 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     6001 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     6061 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     6121 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     6181 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     6241 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     6301 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     6361 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     6421 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     6481 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     6541 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     6601 gcgcacattt ccccgaaaag tgccacctga cgtc
//
