LOCUS       V019708                 8642 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019708
VERSION     V019708
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..3322
                     /label="Tyro3(NM_019392)"
                     /note="Tyro3(NM_019392)"
                     /gene="Tyro3"
     CDS             3329..3394
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3418..4006
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4072..4120
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4322..4750
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4764..5093
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5160..5954
                     /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    6130..6251
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6300..6316)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6324..6340
                     /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(6348..6378)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6393..6414
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6702..7290)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7461..8321)
                     /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(8322..8426)
                     /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 ccatggcgct gaggcggagc atggggtggc cggggctccg
      721 gccgctgctg ctggcgggac tggcttctct gctgctcccc gggtctgcgg ccgcaggcct
      781 gaagctcatg ggcgccccag tgaagatgac cgtgtctcag gggcagccag tgaagctcaa
      841 ctgcagcgtg gaggggatgg aggaccctga catccactgg atgaaggatg gcaccgtggt
      901 ccagaatgca agccaggtgt ccatctccat cagcgagcac agctggattg gcttactcag
      961 cctaaagtca gtggaacggt ctgatgctgg cctgtactgg tgccaggtga aggatgggga
     1021 ggaaaccaag atttctcagt cagtatggct cactgtcgaa ggtgtgccat tcttcacagt
     1081 ggaaccaaaa gatctggcgg tgccacccaa tgcccctttt cagctgtctt gtgaggctgt
     1141 gggtcctcca gaacccgtaa ccatttactg gtggagagga ctcactaagg ttgggggacc
     1201 tgctccctct ccctctgttt taaatgtgac aggagtgacc cagcgcacag agttttcttg
     1261 tgaagcccgc aacataaaag gcctggccac ttcccgacca gccattgttc gccttcaagc
     1321 accgcctgca gctcctttca acaccacagt aacaacgatc tccagctaca acgctagcgt
     1381 ggcctgggtg ccaggtgctg acggcctagc tctgctgcat tcctgtactg tacaggtggc
     1441 acacgcccca ggagaatggg aggcccttgc tgttgtggtt cctgtgccac cttttacctg
     1501 cctgcttcgg aacttggccc ctgccaccaa ctacagcctt agggtgcgct gtgccaatgc
     1561 cttgggccct tctccctacg gcgactgggt gccctttcag acaaagggcc tagcgccagc
     1621 cagagctcct cagaatttcc atgccattcg taccgactca ggccttatcc tggaatggga
     1681 agaagtgatt cctgaggacc ctggggaagg ccccctagga ccttataagc tgtcctgggt
     1741 ccaagaaaat ggaacccagg atgagctgat ggtggaaggg accagggcca atctgaccga
     1801 ctgggatccc cagaaggacc tgattttgcg tgtgtgtgcc tccaatgcaa ttggtgatgg
     1861 gccctggagt cagccactgg tggtgtcttc tcatgaccat gcagggaggc agggccctcc
     1921 ccacagccgc acatcctggg tgcctgtggt cctgggcgtg ctcaccgccc tgatcacagc
     1981 tgctgccttg gccctcatcc tgcttcggaa gagacgcaag gagacgcgtt tcgggcaagc
     2041 ctttgacagt gtcatggccc gaggggagcc agctgtacac ttccgggcag cccgatcttt
     2101 caatcgagaa aggcctgaac gcattgaggc cacattggat agcctgggca tcagcgatga
     2161 attgaaggaa aagctggagg atgtcctcat tccagagcag cagttcaccc tcggtcggat
     2221 gttgggcaaa ggagagtttg gatcagtgcg ggaagcccag ctaaagcagg aagatggctc
     2281 cttcgtgaaa gtggcagtga agatgctgaa agctgacatc attgcctcaa gcgacataga
     2341 agagttcctc cgggaagcag cttgcatgaa ggagtttgac catccacacg tggccaagct
     2401 tgttggggtg agcctccgga gcagggctaa aggtcgtctc cccattccca tggtcatcct
     2461 gcccttcatg aaacatggag acttgcacgc ctttctgctc gcctcccgaa tcggggagaa
     2521 cccttttaac ctgcccctcc agaccctggt ccggttcatg gtggacattg cctgtggcat
     2581 ggagtacctg agctcccgga acttcatcca ccgagaccta gcagctcgga attgcatgct
     2641 ggccgaggac atgacagtgt gtgtggctga ttttggactc tctcggaaaa tctatagcgg
     2701 ggactattat cgtcagggct gtgcctccaa attgcccgtc aagtggctgg ccctggagag
     2761 cttggctgac aacttgtata ctgtacacag tgatgtgtgg gccttcgggg tgaccatgtg
     2821 ggagatcatg actcgtgggc agacgccata tgctggcatt gaaaatgccg agatttacaa
     2881 ctacctcatc ggcgggaacc gcctgaagca gcctccggag tgcatggagg aagtgtatga
     2941 tctcatgtac cagtgctgga gcgccgaccc caagcagcgc ccaagcttca cgtgtctgcg
     3001 aatggaactg gagaacattc tgggccacct gtctgtgctg tccaccagcc aggacccctt
     3061 gtacatcaac attgagagag ctgagcagcc tactgagagt ggcagccctg aggtccactg
     3121 tggagagcga tccagcagcg aggcagggga cggcagtggc gtgggggcag taggtggcat
     3181 ccccagtgac tctcggtaca tcttcagccc cggagggcta tccgagtcac cagggcagct
     3241 ggagcagcag ccagaaagcc ccctcaatga gaaccagagg ctgttgttgc tgcagcaagg
     3301 gctactgcct cacagtagct gtctcgagga ctacaaagac catgacggtg attataaaga
     3361 tcatgacatc gactacaagg atgacgatga caagtagtga gggcccgata tctcgacaat
     3421 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     3481 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     3541 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     3601 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     3661 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     3721 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     3781 ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc
     3841 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     3901 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     3961 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg aaacggggga
     4021 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     4081 aaagacagaa taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
     4141 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
     4201 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
     4261 gtcggggcgg caggccctgc catagcagat ctgcgcagct ggggctctag ggggtatccc
     4321 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     4381 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     4441 acgttcgccg gctttccccg tcaagctcta aatcggggca tccctttagg gttccgattt
     4501 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     4561 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     4621 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     4681 taagggattt tggggatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     4741 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     4801 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     4861 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4921 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     4981 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     5041 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     5101 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
     5161 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     5221 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     5281 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     5341 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     5401 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     5461 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     5521 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     5581 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     5641 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     5701 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     5761 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     5821 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     5881 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     5941 acgagttctt ctgagcggga ctctggggtt cgcgaaatga ccgaccaagc gacgcccaac
     6001 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     6061 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     6121 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     6181 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     6241 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     6301 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     6361 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     6421 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     6481 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     6541 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     6601 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     6661 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     6721 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     6781 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     6841 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc
     6901 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     6961 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     7021 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     7081 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     7141 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     7201 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     7261 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     7321 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     7381 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     7441 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     7501 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     7561 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     7621 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     7681 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     7741 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     7801 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     7861 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     7921 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     7981 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     8041 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     8101 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     8161 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     8221 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     8281 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     8341 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     8401 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
     8461 ggatcgggag atctcccgat cccctatggt cgactctcag tacaatctgc tctgatgccg
     8521 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga
     8581 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta
     8641 gg
//
