LOCUS       V020534                 7052 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020534
VERSION     V020534
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             683..1732
                     /label="CXCR1(NM_000634)"
                     /note="CXCR1(NM_000634)"
                     /gene="CXCR1"
     CDS             1739..1804
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1828..2416
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2482..2530
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2732..3160
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3174..3503
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3570..4364
                     /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    4540..4661
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4710..4726)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4734..4750
                     /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(4758..4788)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4803..4824
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5112..5700)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5871..6731)
                     /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(6732..6836)
                     /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 gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccgcca ccatgtcaaa tattacagat ccacagatgt gggattttga
      721 tgatctaaat ttcactggca tgccacctgc agatgaagat tacagcccct gtatgctaga
      781 aactgagaca ctcaacaagt atgttgtgat catcgcctat gccctagtgt tcctgctgag
      841 cctgctggga aactccctgg tgatgctggt catcttatac agcagggtcg gccgctccgt
      901 cactgatgtc tacctgctga acctggcctt ggccgaccta ctctttgccc tgaccttgcc
      961 catctgggcc gcctccaagg tgaatggctg gatttttggc acattcctgt gcaaggtggt
     1021 ctcactcctg aaggaagtca acttctacag tggcatcctg ctgttggcct gcatcagtgt
     1081 ggaccgttac ctggccattg tccatgccac acgcacactg acccagaagc gtcacttggt
     1141 caagtttgtt tgtcttggct gctggggact gtctatgaat ctgtccctgc ccttcttcct
     1201 tttccgccag gcttaccatc caaacaattc cagtccagtt tgctatgagg tcctgggaaa
     1261 tgacacagca aaatggcgga tggtgttgcg gatcctgcct cacacctttg gcttcatcgt
     1321 gccgctgttt gtcatgctgt tctgctatgg attcaccctg cgtacactgt ttaaggccca
     1381 catggggcag aagcaccgag ccatgagggt catctttgct gtcgtcctca tcttcctgct
     1441 ttgctggctg ccctacaacc tggtcctgct ggcagacacc ctcatgagga cccaggtgat
     1501 ccaggagagc tgtgagcgcc gcaacaacat cggccgggcc ctggatgcca ctgagattct
     1561 gggatttctc catagctgcc tcaaccccat catctacgcc ttcatcggcc aaaattttcg
     1621 ccatggattc ctcaagatcc tggctatgca tggcctggtc agcaaggagt tcttggcacg
     1681 tcatcgtgtt acctcctaca cttcttcgtc tgtcaatgtc tcttccaacc tcctcgagga
     1741 ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg atgacgatga
     1801 caagtagtga gggcccgata tctcgacaat caacctctgg attacaaaat ttgtgaaaga
     1861 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     1921 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     1981 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     2041 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     2101 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     2161 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     2221 aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
     2281 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     2341 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     2401 tgggccgcct ccccgcctgg aaacggggga ggctaactga aacacggaag gagacaatac
     2461 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac gggtgttggg
     2521 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     2581 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     2641 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcagat
     2701 ctgcgcagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     2761 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     2821 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     2881 aatcggggca tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     2941 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     3001 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     3061 aaccctatct cggtctattc ttttgattta taagggattt tggggatttc ggcctattgg
     3121 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     3181 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3241 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3301 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     3361 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     3421 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     3481 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     3541 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     3601 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     3661 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     3721 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     3781 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     3841 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     3901 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     3961 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     4021 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     4081 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     4141 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     4201 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     4261 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     4321 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     4381 cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     4441 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     4501 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     4561 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     4621 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     4681 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     4741 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     4801 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     4861 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     4921 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     4981 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     5041 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     5101 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     5161 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     5221 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     5281 ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag
     5341 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     5401 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     5461 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     5521 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     5581 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     5641 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     5701 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     5761 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     5821 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     5881 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     5941 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     6001 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     6061 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     6121 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     6181 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     6241 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     6301 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     6361 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     6421 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     6481 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     6541 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     6601 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     6661 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     6721 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     6781 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     6841 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
     6901 cgactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
     6961 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
     7021 tgaccgacaa ttgcatgaag aatctgctta gg
//
