LOCUS       V038068                 8742 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V038068
VERSION     V038068
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..3422
                     /label="Itih3(NM_008407)"
                     /note="Itih3(NM_008407)"
                     /gene="Itih3"
     CDS             3429..3494
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3518..4106
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4172..4220
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4422..4850
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4864..5193
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5260..6054
                     /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    6230..6351
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6400..6416)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6424..6440
                     /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(6448..6478)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6493..6514
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6802..7390)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7561..8421)
                     /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(8422..8526)
                     /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 ctcgaattcg ccaccatgag gactatgtgg tggccctgcc
      781 ttgtcttggc tctgctctct ggcttggaga cctctggctt tccgagaagt cccctccagc
      841 tgcttgggaa acggagcctc ccagaggggg tggtggatgg cattgaggtt tacagcacca
      901 agatcagctg caaggtgacc tcccgctttg ctcacaatgt tgtcaccaca agggctgtca
      961 accgtgcaga cacagccaag gaggtgtcct ttgatgtgga actgcccaag acagccttca
     1021 tcaccaactt caccttgacc attgatggtg tcacctaccc cgggaacgtc aaggagaagg
     1081 aagttgccca gaaacaatat gaaaaggctg tgtcccaggg caagacagcc ggtttggtca
     1141 aggcctctgg gaggaaactg gagaaattca cagtgtctgt caacgtggct gcaggcagca
     1201 aagtcacctt tgagctaacc tatgaagaac tgctcaagag aaacaagggc aagtacgaga
     1261 tgtaccttaa ggtccagccc aaacaactgg tcagacactt tgagattgat gcacacatct
     1321 tcgagccaca gggcattagc atgctagatg ctgaagcctc attcatcact aatgacctcc
     1381 tgggaagcgc cctcaccaag tccttctcag gcaagaaggg gcacgtgtct ttcaagccca
     1441 gcttagacca gcagcgttca tgcccgacct gtacagactc cctcctcaac ggggacttca
     1501 ccattgtcta cgatgtgaac agagagtctc cagggaatgt gcagatagtc aatggctact
     1561 ttgtgcactt ctttgcaccc caaggccttc cagtggtgcc taagaacata gtctttgtga
     1621 ttgatgtcag cggctccatg tctggtcgga aaatccagca gaccagggaa gcccttctta
     1681 aaatcctgga tgacgtgaaa gaagatgact acctgaattt cattctcttc agcaccgatg
     1741 tgaccacctg gaaagatcac ctagttcaag ccacccctgc aaacctcaag gaggccaaga
     1801 catttgtgaa gaacatccat gatcaaagca tgaccaacat caatgatggg ctgctgaagg
     1861 gtattgaaat gctgaacaag gcccgagagg accacactgt cccagagagg agcacctcca
     1921 tcatcatcat gttgacagac ggggatgcca atactggcga gagcagacca gaaaagatcc
     1981 aggaaaacgt ccggaatgcc atcgggggca agttcccttt gtataacctg ggttttggca
     2041 acaatctgaa ttataatttc ctggagactc tggccctgga gaaccatggg cttgcccggc
     2101 gcatttatga agattctgat gccaacttgc agctgcaggg cttctatgaa gaggtagcta
     2161 atccactgct gactaatgtg gaggtggaat accccgagaa cgccatccta gacctcacca
     2221 ggaacagtta cccccacttc tacgatggct ctgagattgt tgtagcaggg cgcctggtgg
     2281 acaggaacat ggacaacttc aaagcagatg tgaagggcca tggggccttg aatgacctga
     2341 ccttcacaga ggaggtagac atggaggaaa tggatgcagc cctgaaggag caaggctaca
     2401 tttttgggga ctacattgag cgactctggg cctacctcac tattgagcag ctactggaga
     2461 aacgcaagaa tgccaaaggg gatgagaagg agaacatcac agcggaggcc ctggatctgt
     2521 ccctcaagta ccactttgta accccactga cctccatggt ggtgaccaag cctgaggaca
     2581 atgaagacca gacatccatt gctgacaagc ctggggaaga agccatcgct gagaccacga
     2641 ccatgtcctt cttgaccact cagcagtcca gtcagagccc ctactattat gtggatgggg
     2701 acccccactt tatcatccaa atcccgggaa aaaacgacag catctgcttc aacatcgacg
     2761 agaagcctgg aaccgtgctg cgcctgattc aggacccagt cacaggtatc actgtgaccg
     2821 gacagatcat cggagataag cgaagtaacg cttcttccag gacagggaaa acttattttg
     2881 gcaaactggg cattaccaat gcttggatgg acttccgggt tgaggtgacc acggagaaaa
     2941 tcatcctagg gactggggcc gagctgagca cgttcagttg gctagacaca atcacggtca
     3001 cacagactgg gctgtctgta acaatcaaca ggaaaaagaa tatggtggtg tcctttggag
     3061 atgggattag ctttgtgatc atcctacacc aggtttggaa gaaacatcca gttcaccagg
     3121 acttcctagg gttctacgtg gtggacagtc accggatgtc agcacagaca catgggctgc
     3181 tgggccagtt cttccaaccc tttgacttta aagtgtttgg catccgccca ggctctgacc
     3241 ctacaaagcc agatgccaca atggtggtga agaatcatcg gttgactgtc acaaggggct
     3301 cccagaaaga ttacaggaag gatgccagtg ttggcaccaa ggtcatctgc tggtttgtcc
     3361 ataacaatgg agaaggacta attgatggtg tccataccga ctatattgtt cccagcttgt
     3421 tcggatccga ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg
     3481 atgacgatga caagtagtga gggcccgata tctcgacaat caacctctgg attacaaaat
     3541 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     3601 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     3661 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     3721 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     3781 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     3841 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     3901 gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct
     3961 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     4021 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     4081 gatctccctt tgggccgcct ccccgcctgg aaacggggga ggctaactga aacacggaag
     4141 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac
     4201 gggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     4261 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     4321 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     4381 catagcagat ctgcgcagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     4441 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     4501 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     4561 tcaagctcta aatcggggca tccctttagg gttccgattt agtgctttac ggcacctcga
     4621 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     4681 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     4741 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tggggatttc
     4801 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     4861 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4921 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     4981 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     5041 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     5101 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     5161 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     5221 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     5281 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     5341 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     5401 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     5461 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     5521 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     5581 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     5641 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     5701 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     5761 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     5821 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     5881 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     5941 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     6001 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     6061 ctctggggtt cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     6121 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     6181 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     6241 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     6301 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     6361 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     6421 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     6481 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     6541 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     6601 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     6661 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     6721 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     6781 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     6841 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     6901 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     6961 cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag
     7021 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     7081 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     7141 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     7201 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     7261 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     7321 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     7381 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     7441 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     7501 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     7561 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     7621 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     7681 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     7741 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     7801 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     7861 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     7921 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     7981 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     8041 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     8101 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     8161 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     8221 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     8281 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     8341 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     8401 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     8461 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     8521 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat
     8581 cccctatggt cgactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg
     8641 ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca
     8701 aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gg
//
