LOCUS       V035035                 7692 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035035
VERSION     V035035
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             762..2372
                     /label="TYRP1(NM_000550)"
                     /note="TYRP1(NM_000550)"
                     /gene="TYRP1"
     CDS             2379..2444
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2468..3056
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3122..3170
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3372..3800
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3814..4143
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4210..5004
                     /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    5180..5301
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5350..5366)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5374..5390
                     /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(5398..5428)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5443..5464
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5752..6340)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6511..7371)
                     /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(7372..7476)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcctcgagg aattcgccac catgagtgct cctaaactcc
      781 tctctctggg ctgtatcttc ttccccttgc tactttttca gcaggcccgg gctcaattcc
      841 caagacagtg tgccactgtt gaggctttga gaagtggtat gtgttgccca gacctgtccc
      901 ctgtgtctgg gcctgggaca gaccgctgtg gctcatcatc agggaggggc agatgtgagg
      961 cagtgactgc agactcccgg ccccacagcc ctcagtatcc ccatgatggc agagatgatc
     1021 gggaggtctg gcccttgcgc ttcttcaata ggacatgtca ctgcaacggc aatttctcag
     1081 gacacaactg tgggacgtgc cgtcctggct ggagaggagc tgcctgtgac cagagggttc
     1141 tcatagtcag gagaaatctt ctggacttaa gtaaagaaga aaagaaccac tttgtccggg
     1201 ccctggatat ggcaaagcgc acaactcacc ctttatttgt cattgccacc aggagatcag
     1261 aagaaatact ggggccagat ggcaacacgc cacaatttga gaacatttcc atttataact
     1321 actttgtttg gacacactat tactcagtca aaaagacttt ccttggggta ggacaggaaa
     1381 gctttggtga agtggatttc tctcatgagg gaccagcttt tctcacatgg cacaggtacc
     1441 acctcctgcg tctggagaaa gacatgcagg aaatgttgca agagccttct ttctcccttc
     1501 cttactggaa ttttgcaacg gggaaaaatg tctgtgatat ctgcacggat gacttgatgg
     1561 gatccagaag caactttgat tccactctaa taagcccaaa ctctgtcttt tctcaatggc
     1621 gagtggtctg tgactccttg gaagattatg ataccctggg aacactttgt aacagcaccg
     1681 aggatgggcc aattaggaga aatccagctg gaaatgtggc cagaccaatg gtgcaacgtc
     1741 ttcctgaacc acaggatgtc gctcagtgct tggaagttgg tttatttgac acgcctcctt
     1801 tttattccaa ctctacaaac agtttccgaa acacagtgga aggttacagt gaccccacgg
     1861 gaaagtatga ccctgctgtt cgaagtcttc acaatttggc tcatctattc ctgaatggaa
     1921 cagggggaca aacccatttg tctccaaatg atcctatttt tgtcctcctg cacaccttca
     1981 cagatgcagt ctttgatgaa tggctgagga gatacaatgc tgatatatcc acatttccat
     2041 tggaaaatgc ccctattgga cataatagac aatacaacat ggtgccattc tggcccccag
     2101 tcaccaacac agaaatgttt gttactgctc cagacaacct gggatacact tatgaaattc
     2161 aatggccaag tcgggagttt agtgtacctg agataattgc catagcagta gttggcgctt
     2221 tgttactggt tgcactcatt tttgggactg cttcttatct gattcgtgcc agacgcagta
     2281 tggatgaagc taaccagcct ctcctcactg atcagtatca atgctatgct gaagaatatg
     2341 aaaaactcca gaatcctaat cagtctgtgg tcggatccga ctacaaagac catgacggtg
     2401 attataaaga tcatgacatc gactacaagg atgacgatga caagtagtga gggcccgata
     2461 tctcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     2521 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     2581 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     2641 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     2701 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     2761 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     2821 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg
     2881 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     2941 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     3001 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg
     3061 aaacggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     3121 acggcaataa aaagacagaa taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg
     3181 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     3241 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     3301 cgcagccaac gtcggggcgg caggccctgc catagcagat ctgcgcagct ggggctctag
     3361 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     3421 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     3481 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcggggca tccctttagg
     3541 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     3601 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     3661 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     3721 ttttgattta taagggattt tggggatttc ggcctattgg ttaaaaaatg agctgattta
     3781 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     3841 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     3901 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3961 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     4021 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     4081 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     4141 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     4201 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     4261 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     4321 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     4381 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     4441 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     4501 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     4561 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     4621 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     4681 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     4741 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     4801 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     4861 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     4921 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     4981 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgcgaaatga ccgaccaagc
     5041 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     5101 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     5161 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     5221 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     5281 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     5341 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     5401 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     5461 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     5521 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     5581 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     5641 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     5701 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     5761 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     5821 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     5881 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     5941 ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     6001 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     6061 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     6121 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     6181 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     6241 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     6301 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     6361 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     6421 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     6481 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     6541 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     6601 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     6661 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     6721 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     6781 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     6841 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     6901 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     6961 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     7021 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     7081 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     7141 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     7201 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     7261 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     7321 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     7381 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     7441 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     7501 tgacgtcgac ggatcgggag atctcccgat cccctatggt cgactctcag tacaatctgc
     7561 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
     7621 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
     7681 aatctgctta gg
//
