LOCUS       V047440                10749 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V047440
VERSION     V047440
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..5429
                     /label="Rere(NM_001085492)"
                     /note="Rere(NM_001085492)"
                     /gene="Rere"
     CDS             5436..5501
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    5525..6113
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    6179..6227
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      6429..6857
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6871..7200
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             7267..8061
                     /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    8237..8358
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(8407..8423)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    8431..8447
                     /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(8455..8485)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    8500..8521
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8809..9397)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(9568..10428)
                     /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(10429..10533)
                     /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 ccaccatgac agctgacaaa gataaagaca
      781 aagacaaaga gaaggaccgg gaccgggatc gggaccgaga aagagataaa agagacaaag
      841 caagagagag cgagaatgcc aggcctcgca ggagctgtac cttggaggga ggagccaaaa
      901 actatgctga gagcgatcac agcgaggacg aagacaacga caacaacagc gccaccacgg
      961 aggagtccaa caagaagagc agaaagaaac ctccgaaaaa aaagtcccgc tacgagagga
     1021 cagacactgg tgagataact tcttacatca ccgaggacga cgttgtctac agaccgggag
     1081 actgtgtgta tatcgagagt cggcggccaa acacaccgta tttcatctgc agcatccaag
     1141 acttcaaact ggtccacagc tcccaggcct gttgcagatc tccagctcct gctttctgtg
     1201 accccccagc atgctctctg ccggtggcac cacagccacc acagcacctc tctgaagccg
     1261 ggagagggcc tggagggagt aagagggacc atctactcat gaacgtcaaa tggtactacc
     1321 gtcagtctga agtcccagat tctgtctatc agcatttggt tcaggatcgg cataatgaaa
     1381 acgactctgg aagagagctt gtcatcacag atccagtcat caagaatcgg gaactcttca
     1441 tttctgatta tgttgacacc taccatgctg ctgccctaag agggaagtgt aacatctccc
     1501 atttttctga catatttgct gctcgagagt ttaaagcccg agtggactca tttttctaca
     1561 tattaggata caaccctgag acaaggcggc tgaatagtac ccaaggggaa attcgagttg
     1621 gccctagcca ccaggccaaa cttccagatc tgcaaccatt tccttctcca gatggtgata
     1681 ctgtgactca gcatgaggaa cttgtctgga tgcctggagt cagtgactgt gacctcctca
     1741 tgtacttgag ggcagcaagg agcatggcag catttgcagg gatgtgtgat ggaggctcca
     1801 cagaggatgg ctgtgttgca gcatctcggg atgacaccac tctcaatgca ctgaacacac
     1861 tacatgaaag cagttatgat gccggcaaag ccctgcagcg cctggtgaag aagcctgtgc
     1921 ccaagctgat cgagaagtgc tggacagagg atgaagtgaa acgcttcgtt aaggggctcc
     1981 gacaatacgg caagaacttc ttcagaatca gaaaggagct gcttcccagt aaggaaaccg
     2041 gggaactgat taccttctat tactactgga agaaaacacc agaggcggcc agctcccgtg
     2101 cccaccgccg ccaccgcagg caggcggtgt tcagaagaat taagactcgc acagcgtcca
     2161 cccctgtcaa cacgccctcc agaccacctt ccagtgaatt cttggacctg agttcagcta
     2221 gtgaggatga ctttgacagt gaggacagcg aacaggagct gaaggggtac gcctgccgtc
     2281 actgcttcac caccacctcc aaagactggc accacggggg ccgggaaaac atcctactgt
     2341 gcacggactg tcgcatccac ttcaagaagt acggtgagct gccacccatc gagaagcccg
     2401 tggacccccc gcccttcatg ttcaagcctg tcaaggagga ggacgatggg ctcagtggga
     2461 agcatagcat gaggacgcgg cggagccggg gctcgatgtc tacactacgc agtggtcgga
     2521 agaagcagcc caccagtcct gatggtcgtg cctcacccat caatgaagac atccgatcca
     2581 gtggccggaa ctcccccagt gctgctagca cttccagcaa tgacagtaaa gcagagacag
     2641 tgaagaagtc agccaagaag gtgaaggaag aggctgcatc ccctctgaag agcaccaagc
     2701 gccagcggga gaaggtggcc tctgacacag aagacactga taggatcacc tccaaaaaga
     2761 caaagaccca ggagattagc cgtcccaact ctccctctga aggagagggg gagagttcgg
     2821 acagtcgcag tgtcaacgat gagggcagca gtgacccgaa agacattgac caggataatc
     2881 gcagcacgtc tcccagtatc cccagccctc aggataatga gagtgactcg gactcatcag
     2941 cacagcagca gatgttgcag gcccagcccc cagccttgca agctcccagt ggtgctgcct
     3001 cggctccctc cacagcccct ccaggaacac cccagcttcc cacccaagga cccacaccct
     3061 ctgccactgc agtccctcca cagggctccc ctgcaacctc acagccccct aaccagactc
     3121 agtctacagt tgcacctgca gcccacaccc acatccaaca ggcacccacc ctgcaccccc
     3181 ctcggctgcc ctcacctcat cctccactgc agcctatgac tgcaccgcct agccagagct
     3241 ctgcacaacc tcacccccag ccctccttgc acagtcaggg cccgcctggc cctcacagcc
     3301 tacagactgg acccctgtta cagcacccag gacctcctca gccctttgga cttccttccc
     3361 agccctccca aggtcagggc cctctgggcc cctccccggc agcagctcat cctcatagca
     3421 ccatacagct tccagcgtcg cagtcagcgc tgcagcccca acagccccca agggaacagc
     3481 ccttgcctcc tgccccttta gctatgcctc acatcaaacc cccacccacc acaccaattc
     3541 cccagcttcc agcaccacaa gcccacaaac atccgcctca cctctcaggc ccctccccct
     3601 tctctctgaa tgccaatctg ccacccccgc cagccctgaa acccctgagc tcactgtcca
     3661 cacaccaccc tccctcagcc caccctccac ccttacaact catgcctcag agccagccct
     3721 tgccctcttc acctgcccag ccacctgggc tgacccagag ccagagccta ccacctcctg
     3781 ctgcctccca tcctaccact ggcctccacc aggtgccctc ccaatcccca tttccccagc
     3841 accctttcgt acctggaggc ccccctccca tcaccccgcc ctcctgcccc cccacttcca
     3901 cccctcctgc cgggcccagc tcctcatcac agccaccctg ctctgctgct gtgtcttctg
     3961 gaggcagtgt tcctggggcg ccatcttgcc cactccctgc tgtgcaaatc aaggaggagg
     4021 ctctggatga agctgaggag ccagagagcc ctcctccacc ccccaggagc ccatcccctg
     4081 agcccactgt agtggacacc cccagccatg ccagccagtc tgccaggttc tacaaacacc
     4141 tggaccgggg ctacaactcg tgtgcacgga cagacctgta cttcatgcct ctggctggat
     4201 ccaaactagc caagaagagg gaggaggcca ttgagaaggc caagcgggag gctgaacaga
     4261 aagcccgaga ggaacgggag cgggagaaag agaaggaaaa agagcgagaa cgggagagag
     4321 agcgggaacg ggaggcagag cgcgctgcta aggcatccag ctctgcacat gaaggccgtc
     4381 ttagtgaccc tcagctcagt ggcccgggtc acatgcgacc ctccttcgag ccaccaccga
     4441 ccacaatcgc tgctgtgccc ccatatattg ggcctgacac acctgccctc cggacactga
     4501 gcgaatatgc tcggcctcat gtcatgtctc ccaccaaccg caaccacccc ttctacatgc
     4561 cccttaatcc caccgacccc ctgctggcct accacatgcc tggcttgtac aatgttgatc
     4621 ccaccatccg cgagcgggag ctccgagagc gcgagatcag agagcgcgag atccgggagc
     4681 gggaattgcg ggagaggatg aagccaggct ttgaggtgaa gcccccagag ctggaccccc
     4741 tgcacccagc caccaacccc atggagcatt ttgcccggca cagcgccctc accatccctc
     4801 ctgcagctgg cccccacccc tttgcctctt tccacccggg cctcaaccct ttggagcggg
     4861 agagactggc actagcaggt cctcagctgc ggcctgagat gagttaccca gacagactgg
     4921 cagccgagcg catccatgct gagcgtatgg catcactcac cagcgacccc ctggcacgac
     4981 tgcagatgtt taatgtcact ccacaccatc accagcattc acacatccac tcccacctcc
     5041 acctccacca gcaggatcct ctccaccaag gttcagcagg cccagttcac cccctggttg
     5101 accccttgac tgctggccct cacttagctc gcttccccta tcccccgggc accctcccta
     5161 accctctact tggacagccc cctcacgagc acgagatgct gcgccaccca gtttttggca
     5221 ccccctatcc ccgagacctg cctggggcta ttccaccccc catgtcagca gcccaccagc
     5281 ttcaggccat gcatgcccag tcagccgagc tacagagact agccatggag cagcagtggc
     5341 tgcacggaca tccacacatg catggtggcc atctgccaag tcaggaggat tattacagtc
     5401 gactgaagaa agaaggtgac aagcagttag gatccgacta caaagaccat gacggtgatt
     5461 ataaagatca tgacatcgac tacaaggatg acgatgacaa gtagtgaggg cccgatatct
     5521 cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     5581 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     5641 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     5701 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     5761 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     5821 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     5881 gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct
     5941 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     6001 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     6061 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaaa
     6121 cgggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     6181 gcaataaaaa gacagaataa aacgcacggg tgttgggtcg tttgttcata aacgcggggt
     6241 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     6301 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     6361 agccaacgtc ggggcggcag gccctgccat agcagatctg cgcagctggg gctctagggg
     6421 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     6481 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     6541 tctcgccacg ttcgccggct ttccccgtca agctctaaat cggggcatcc ctttagggtt
     6601 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     6661 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     6721 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     6781 tgatttataa gggattttgg ggatttcggc ctattggtta aaaaatgagc tgatttaaca
     6841 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     6901 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     6961 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     7021 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     7081 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     7141 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     7201 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     7261 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     7321 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     7381 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     7441 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     7501 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     7561 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     7621 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     7681 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     7741 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     7801 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     7861 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     7921 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     7981 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     8041 cttcttgacg agttcttctg agcgggactc tggggttcgc gaaatgaccg accaagcgac
     8101 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     8161 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     8221 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     8281 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     8341 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     8401 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     8461 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     8521 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     8581 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     8641 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     8701 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     8761 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     8821 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     8881 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     8941 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     9001 tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     9061 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     9121 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     9181 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     9241 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     9301 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     9361 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     9421 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     9481 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     9541 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     9601 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     9661 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     9721 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     9781 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     9841 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     9901 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     9961 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
    10021 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
    10081 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
    10141 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
    10201 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
    10261 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
    10321 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
    10381 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
    10441 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
    10501 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
    10561 cgtcgacgga tcgggagatc tcccgatccc ctatggtcga ctctcagtac aatctgctct
    10621 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
    10681 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
    10741 ctgcttagg
//
