LOCUS       V023577                 8922 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023577
VERSION     V023577
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             756..3602
                     /label="Rai14(NM_001011947)"
                     /note="Rai14(NM_001011947)"
                     /gene="Rai14"
     CDS             3609..3674
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3698..4286
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4352..4400
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4602..5030
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5044..5373
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5440..6234
                     /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    6410..6531
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6580..6596)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6604..6620
                     /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(6628..6658)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6673..6694
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6982..7570)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7741..8601)
                     /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(8602..8706)
                     /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 ctcggatccg ccaccatgaa gagcctgaaa gcgaagttta
      781 ggaagagcga cacgaacgag tggaacaaga atgatgaccg cctgctgcag gctgtggaga
      841 atggagatgc tgagaaggtg gcctcactgc tgggcaagaa aggggccagt gccaccaagc
      901 atgacagcga gggcaagacc gctttccacc ttgctgctgc aaaaggacac gtggaatgcc
      961 tcaaggtcat ggttacacac ggtgtggatg tgactgccca agactcttcc ggtcacagtg
     1021 ctttacatat cgcagccaag aatggccacc cagagtacat caagaagctt cttcagtaca
     1081 agagcccagc cgaaagtgtt gacaacttgg ggaaaacagc tttgcattat gcagcggcac
     1141 aggggagcct tcaagctgta caagtcctct gcgaacacaa gagccccata aacctcaaag
     1201 acttggatgg gaacataccc ctgctcgtcg caatacaaaa tggccacagt gaggcctgcc
     1261 actttctcct ggaccatggc gcagatgtca attccaggga caaaaatgga agaactgctc
     1321 tcatgctggc gtgcgaaact ggcagtgcta acactgtgga agccttaatt aaaaaggggg
     1381 cagacctgaa ccttgtagat tctcttggac acaatgcctt atattattcc aaactctcag
     1441 aaaatgcagg aattcagagc cttctattat caaaaatctc tcaggatgct gatttaaaga
     1501 ccccaacaaa agcaaagcag ttgagtgatg tgtcttcccc acgatcaata acttctacac
     1561 cactttcagg aaaggaatca gtattttttg ctgaagcacc ctttaaggct gagatcagct
     1621 ccatacaaga aaacaaagac aggctgagcg acagtactgc aggtgctgac agcttattag
     1681 atgtaagttc tgaagctgac caacaagatc ttcttgtcct attgcaagca aaagtcgctt
     1741 cccttacctt acacaataag gagttacaag ataaattgca ggccaaatca cccaaggaaa
     1801 ctgaagccga cctaagcttt cagtctttcc attccaccca gactgaccta gccccatccc
     1861 cgagcaaatc tagtgacatc ccttcctccg atgccaaatc atccccgcct gttgaacatc
     1921 cggcggggac atccaccgct gacagggatg tcataatcca gcagctacaa gataccttgc
     1981 atgatttgca gaagagattg gagacctctg aagcagagaa gaaacagctc caggacgaac
     2041 tccagtcaca aaggacagac acgacatgct tgaacaacac agagatttcc gaaaatggct
     2101 ccgacctcag tcagaaactc aaagacactc agagcaagta cgaagaggcc atgaaagaag
     2161 tcctgagcgt gcagaagcag atgaagctgg gtctcctctc ccacgagagt gccgatggcg
     2221 actcgcgtct ccgagaggtg agggtcactg atgaagacgt tgacgcacta aagcaagacc
     2281 tgcagagagc actggaagag agtaaaaggg acaaggcacg ggtgcaggag ttagagacaa
     2341 agctcgtgga gaaagaaaag gctgaagcga ctaagccgtc ctcagaggtg tgcgaggaga
     2401 tgagaaactc ctactgctct gtcatcgaga atatgaataa ggagaaggca tttttgtttg
     2461 agaaatacca gcaggcccaa gaagaaatca tgaaactaaa ggacacacta aaaagtcaga
     2521 tgccacagga agcccccgat gactctgggg acatgaaaga gaccatgaac aggatggttg
     2581 acgagctcaa taagcaggtg agcgagctgt cacagctcta cagagaagcc caggcagagc
     2641 tggaggatta caggaagagg aagtctctag aagacgcaac cgaatacatt cacagagcgg
     2701 aacacgagag gctgatgcac ctgtcaaacc tgtccaggac caaagcggaa gaatcgctgt
     2761 ccgacatgag gtctcaatat tccaaggtgc tgaacgagtt gactcagctc aagcagctgg
     2821 tggatgcgca caaagagaac tccgtgtcca tcacagagca tctggaagta ataaccacgc
     2881 tgcggacgat ggccaaagag atggaagaga aaacagtcac cctccaagag cacctcgcaa
     2941 gcaaggaagg ggaagtcgca aagctggaga agcagctcgc agaggagaaa gccgccatga
     3001 gcgacgcaat ggtccccaag gcttcctacg agaagctcca ggcctccctg gagagcgaag
     3061 tcaacgcgtt ggcagcaaag ttaaaggagt cggtgaaaga gaaagagaaa gcccattctg
     3121 aggtcgccca ggttcgaagt gaggtctcgc aggcgaaaag ggaaaaggaa aacatccaga
     3181 ctctcttgaa atccaaagag caggaagtaa ctgaacttgt gcagaaattc cagcgagctc
     3241 aggaagaact ggccgggttg aaaagatgct ctgagacctc gtcgaagctg gaggaggaca
     3301 aagacgaaaa gataaatgaa atgaccaagg aggtcctgaa gctgaaggag gccctgaaca
     3361 gcctctcgca gctctcatac tccacaagct cgtccaagag gcagacccag caactggaaa
     3421 tgctacagca gcaggtcaag cagctgcaga accagctggc tgaatgtaag aaacaacacc
     3481 aggaggtcgt atccgtttac agaatgcatc ttctatatgc tgtacagggc caaatggatg
     3541 aagatgtcca gaaagtcttg aagcagatcc tcaccatgtg taaaaaccag tctcagaaga
     3601 agctcgagga ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg
     3661 atgacgatga caagtagtga gggcccgata tctcgacaat caacctctgg attacaaaat
     3721 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     3781 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     3841 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     3901 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     3961 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     4021 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     4081 gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct
     4141 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     4201 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     4261 gatctccctt tgggccgcct ccccgcctgg aaacggggga ggctaactga aacacggaag
     4321 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac
     4381 gggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     4441 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     4501 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     4561 catagcagat ctgcgcagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     4621 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     4681 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     4741 tcaagctcta aatcggggca tccctttagg gttccgattt agtgctttac ggcacctcga
     4801 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     4861 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     4921 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tggggatttc
     4981 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     5041 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     5101 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     5161 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     5221 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     5281 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     5341 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     5401 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     5461 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     5521 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     5581 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     5641 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     5701 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     5761 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     5821 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     5881 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     5941 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     6001 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     6061 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     6121 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     6181 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     6241 ctctggggtt cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     6301 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     6361 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     6421 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     6481 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     6541 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     6601 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     6661 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     6721 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     6781 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     6841 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     6901 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     6961 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     7021 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     7081 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     7141 cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag
     7201 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     7261 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     7321 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     7381 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     7441 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     7501 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     7561 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     7621 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     7681 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     7741 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     7801 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     7861 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     7921 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     7981 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     8041 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     8101 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     8161 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     8221 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     8281 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     8341 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     8401 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     8461 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     8521 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     8581 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     8641 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     8701 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat
     8761 cccctatggt cgactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg
     8821 ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca
     8881 aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gg
//
