LOCUS       V026564                10017 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V026564
VERSION     V026564
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..4697
                     /label="SYNRG(NM_007247)"
                     /note="SYNRG(NM_007247)"
                     /gene="SYNRG"
     CDS             4704..4769
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4793..5381
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5447..5495
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5697..6125
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6139..6468
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6535..7329
                     /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    7505..7626
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7675..7691)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7699..7715
                     /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(7723..7753)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7768..7789
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8077..8665)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8836..9696)
                     /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(9697..9801)
                     /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 ccaccatggc gctgcggcca ggagctggtt
      781 ctggtggcgg cggggccgcg ggagctggcg cggggtccgc cgggggaggc ggcttcatgt
      841 ttcctgttgc aggtgggata agaccccctc aagcaggcct gatgccgatg cagcaacaag
      901 gatttcctat ggtctctgtc atgcagccta atatgcaagg cattatggga atgaattaca
      961 gctctcagat gtcccaagga cctattgcta tgcaggcagg aataccaatg ggaccaatgc
     1021 cagcagcggg aatgccttac ctaggacaag cacccttcct gggcatgcgt cctccaggcc
     1081 cacagtacac tccagacatg cagaagcagt ttgccgaaga gcagcagaaa cgatttgaac
     1141 agcagcaaaa actcttagaa gaagaaagaa aaagacgcca gtttgaagag cagaagcaaa
     1201 agctcagact tttgagcagt gtgaaaccca agacaggaga gaagagtaga gatgatgctt
     1261 tggaagccat aaaaggaaat ttagatgggt tttccagaga tgcaaaaatg caccctactc
     1321 cagcatcgca ccccaagaaa ccaggccctt ccttggagga gaagttccta gtatcttgtg
     1381 atataagtac atctgggcag gaacaaatta aattaaatac ttctgaagtt ggccacaaag
     1441 ccctaggccc aggttccagt aagaagtatc ccagtttaat ggccagtaac ggggttgctg
     1501 tagatggatg tgtaagtggt accaccactg cagaggcaga aaatacttca gatcaaaacc
     1561 tgtcaattga agagagtggt gtgggagtat ttccctcaca ggatcctgct cagcccagaa
     1621 tgcctccttg gatttacaat gagagtttgg ttccagatgc ctataagaaa atcttagaaa
     1681 ccacaatgac tccaactgga atagatactg ccaaactgta tcccattctg atgtcatctg
     1741 ggcttcccag ggaaactctt ggacagatat gggccttagc taatcgaact acacctggca
     1801 aacttacaaa agaagaactt tataccgttc tagccatgat agcggtaaca cagaggggcg
     1861 ttcctgcaat gagtcctgat gctttaaacc agttcccagc agctcctatt ccaactttaa
     1921 gtggcttttc tatgactctg cctacaccgg tgagtcagcc aactgtgata ccttcaggtc
     1981 ctgcgggctc catgcccctc agccttggac agccagtcat gggcattaac cttgttggac
     2041 cagtgggtgg agctgcagcc caggcttcta gtggtttcat accaacctac cctgcaaatc
     2101 aggtagtaaa gccagaagaa gatgacttcc aggattttca agatgcttct aagtcaggat
     2161 cccttgatga ctcattcagt gatttccaag agttgcctgc ttcttcaaaa acaagtaact
     2221 cccagcatgg aaacagtgcc ccttctttgt tgatgccact tcctggaact aaagcattgc
     2281 cttcaatgga caaatatgct gtgtttaaag gaattgcagc tgacaagtcc tctgaaaata
     2341 ctgttccacc tggagatcct ggtgataaat atagtgcttt cagagaactt gaacagacag
     2401 cagagaataa acctttagga gaaagctttg cagaattcag atctgcagga actgatgatg
     2461 gtttcaccga ttttaaaaca gccgatagtg tatcaccact agagccacca acaaaagaca
     2521 aaacttttcc accatccttc ccctcaggaa ctatacaaca gaaacaacaa acacaagtga
     2581 aaaaccctct gaacttagca gacctagata tgttttcctc agttaattgc agcagcgaga
     2641 aaccattgtc tttttcagct gtgtttagca catcaaaatc agtttctaca ccacagtcaa
     2701 caggttctgc tgctactatg acagcattgg cagcaacaaa aacttctagt ttggctgatg
     2761 attttggaga attcagcctt tttggggaat attctggtct agcacctgtt ggggagcagg
     2821 atgactttgc agattttatg gctttcagta atagctctat ttcatctgag caaaagccgg
     2881 atgacaaata tgatgccctt aaagaggaag ccagtcctgt tcctctaacc agcaacgtgg
     2941 gcagcacagt gaagggtgga caaaactcga ctgctgcgtc taccaagtac gatgtcttca
     3001 gacaactttc tctggaaggg tctggactag gtgttgaaga cctgaaagat aacactcctt
     3061 caggaaaaag tgatgatgat tttgctgact tccactccag taaattttct tccataaact
     3121 cggacaaatc cctgggagag aaagcagtgg ctttcagaca caccaaagaa gactctgcat
     3181 cagtgaagtc cttagatctc ccttccattg gtggcagcag tgttggcaag gaggactctg
     3241 aagatgcact ctctgttcag tttgacatga aattggctga tgtgggagga gatcttaagc
     3301 atgtcatgtc tgatagctct ttggatttac caacagttag tggccagcat cctcctgctg
     3361 cagatataga ggacttaaaa tatgctgctt ttggaagcta cagtagcaat tttgcagtga
     3421 gcacacttac aagctatgac tggtcagaca gggatgatgc aactcagggc agaaaactct
     3481 ctccatttgt cctctcagca ggaagtggat ccccctcagc cacctcaatt cttcaaaaga
     3541 aagagacttc atttggcagt tctgaaaaca tcaccatgac atctctctcc aaagtaacga
     3601 cctttgtaag tgaagatgct cttccagaga ccaccttccc agctcttgcc agttttaaag
     3661 acacgattcc tcagaccagt gagcaaaagg aatatgaaaa cagagactat aaagatttca
     3721 caaaacagga cctgcctacg gctgaacgga gccaggaggc cacgtgtccc agcccagcgt
     3781 ccagtggtgc ctctcaagaa accccgaacg aatgttcgga tgactttgga gagtttcaaa
     3841 gtgaaaagcc caaaatcagc aaatttgact tcttagtagc cacttcacaa agcaaaatga
     3901 aatccagtga agaaatgatc aaaagtgagc tggcaacctt tgacctttct gttcaaggat
     3961 cacacaagag gagtttgagc cttggtgata aagaaataag ccgttcttct ccttctccag
     4021 ctttggagca gcctttcaga gaccgttcca atactctgaa tgagaagccc gccctgcccg
     4081 tcatccgaga caagtacaaa gacctgacgg gagaggtgga ggaaaatgag agatatgcat
     4141 atgaatggca gagatgcctg gggagtgccc tgaatgtcat taagaaggca aatgatacct
     4201 taaatggaat cagtagtagt tctgtttgca cagaagtaat tcagtcagct caaggcatgg
     4261 aatatttatt aggtgttgtt gaagtgtaca gggtaaccaa gcgtgtggag ctggggataa
     4321 aagccactgc agtgtgcagt gagaaactcc agcagttgct gaaggacatc gataaagtat
     4381 ggaataacct aatcggcttc atgtcactcg ccacactcac accagatgaa aactcgctgg
     4441 atttttcctc ctgtatgtta cggcctggga ttaaaaatgc tcaggagctt gcctgtggag
     4501 tgtgcctctt gaatgtggac tcgaggagcc ggaaagaaga gaagcctgca gaagaacatc
     4561 ctaaaaaagc attcaactca gaaacagaca gtttcaagct ggcctatgga gggcaccagt
     4621 atcacgccag ctgtgccaac ttctggatca actgtgtcga accaaagcct cctggcctcg
     4681 tcctgcctga cctgctcgga tccgactaca aagaccatga cggtgattat aaagatcatg
     4741 acatcgacta caaggatgac gatgacaagt agtgagggcc cgatatctcg acaatcaacc
     4801 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     4861 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     4921 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     4981 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     5041 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     5101 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     5161 tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt
     5221 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     5281 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     5341 ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggaaacg ggggaggcta
     5401 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga
     5461 cagaataaaa cgcacgggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     5521 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     5581 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     5641 ggcggcaggc cctgccatag cagatctgcg cagctggggc tctagggggt atccccacgc
     5701 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     5761 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     5821 cgccggcttt ccccgtcaag ctctaaatcg gggcatccct ttagggttcc gatttagtgc
     5881 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     5941 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     6001 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     6061 gattttgggg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     6121 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     6181 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     6241 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     6301 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     6361 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     6421 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     6481 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     6541 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     6601 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     6661 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     6721 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     6781 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     6841 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     6901 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     6961 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     7021 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     7081 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     7141 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     7201 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     7261 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     7321 ttcttctgag cgggactctg gggttcgcga aatgaccgac caagcgacgc ccaacctgcc
     7381 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     7441 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     7501 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     7561 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     7621 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     7681 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     7741 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     7801 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     7861 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     7921 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     7981 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     8041 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     8101 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     8161 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     8221 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg
     8281 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     8341 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     8401 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     8461 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     8521 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     8581 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     8641 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     8701 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     8761 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     8821 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     8881 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     8941 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     9001 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     9061 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     9121 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     9181 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     9241 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     9301 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     9361 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     9421 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     9481 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     9541 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     9601 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     9661 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     9721 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     9781 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
     9841 gggagatctc ccgatcccct atggtcgact ctcagtacaa tctgctctga tgccgcatag
     9901 ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa
     9961 atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagg
//
