LOCUS       V026449                 6812 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V026449
VERSION     V026449
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             941..2302
                     /label="GABPA(NM_002040)"
                     /note="GABPA(NM_002040)"
                     /gene="GABPA"
     CDS             2309..2374
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2412..2636
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2682..3110
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3124..3453
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3520..4314
                     /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    4488..4609
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4658..4674)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4682..4698
                     /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(4706..4736)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4751..4772
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5060..5645)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5816..6676)
                     /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(6677..6781)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccgccacc atgactaaaa gagaagcaga
      961 ggagctgata gaaattgaga ttgatggaac agagaaagca gagtgcacag aagaaagcat
     1021 tgtagaacaa acctacgcgc cagctgaatg tgtaagccag gccatagaca tcaatgaacc
     1081 aataggcaat ttaaagaaac tgctagaacc aagactacag tgttctttgg atgctcatga
     1141 aatttgtctg caagatatcc agctggatcc agaacgaagt ttatttgacc aaggagtaaa
     1201 aacagatgga actgtacagc ttagtgtaca ggtaatttct taccaaggaa ttgaaccaaa
     1261 gttaaacatc cttgaaattg ttaaacctgc ggacactgtt gaggttgtta ttgatccaga
     1321 tgcccaccat gctgaatcag aagcacatct tgttgaagaa gctcaagtga taactcttga
     1381 tggcacaaaa cacatcacaa ccatttcaga tgaaacttca gaacaagtga caagatgggc
     1441 tgctgcactg gaaggctata ggaaagaaca agaacgcctt gggataccct atgatcccat
     1501 acagtggtcc acagaccaag tcctgcattg ggtggtttgg gtaatgaagg aattcagcat
     1561 gaccgatata gacctcacca cactcaacat ttcggggaga gaattatgta gtctcaacca
     1621 agaagatttt tttcagcggg ttcctcgggg agaaattctc tggagtcatc tggaacttct
     1681 ccgaaaatat gtattggcaa gtcaagaaca acagatgaat gaaatagtta caattgatca
     1741 acctgtgcaa attattccag catcagtgca atctgctaca cctactacca ttaaagttat
     1801 aaatagtagt gcgaaagcag ccaaagtaca aagagcgccg aggatttcag gagaagatag
     1861 aagctcacct gggaacagaa caggaaacaa tggccaaatc caactatggc agtttttgct
     1921 agaacttctt actgataagg acgctcgaga ctgcatttct tgggttggtg atgaaggtga
     1981 atttaagcta aatcagcctg aactggttgc acagaaatgg ggacagcgta aaaataagcc
     2041 tacgatgaac tatgagaaac tcagtcgtgc attaagatat tattacgatg gggacatgat
     2101 ttgtaaagtt caaggcaaga gatttgtgta caagtttgtc tgtgacttga agactcttat
     2161 tggatacagt gcagcggagt tgaaccgttt ggtcacagaa tgtgaacaga agaaacttgc
     2221 aaagatgcag ctccatggaa ttgcccagcc agtcacagca gtagctctgg ctactgcttc
     2281 tctgcaaacg gaaaaggata atctcgagga ctacaaagac catgacggtg attataaaga
     2341 tcatgacatc gactacaagg atgacgatga caagtagtga gggcccgttt aaacccgctg
     2401 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     2461 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     2521 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     2581 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     2641 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     2701 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     2761 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     2821 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     2881 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     2941 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     3001 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     3061 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     3121 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3181 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     3241 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     3301 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     3361 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     3421 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     3481 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     3541 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     3601 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     3661 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     3721 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     3781 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     3841 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     3901 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     3961 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     4021 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     4081 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     4141 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     4201 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     4261 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     4321 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     4381 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     4441 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg
     4501 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     4561 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
     4621 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     4681 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     4741 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     4801 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     4861 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     4921 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     4981 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     5041 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     5101 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     5161 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     5221 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     5281 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     5341 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     5401 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     5461 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     5521 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     5581 ccaccgctgg tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     5641 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     5701 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     5761 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     5821 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     5881 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     5941 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     6001 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     6061 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     6121 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     6181 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     6241 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     6301 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     6361 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     6421 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     6481 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     6541 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     6601 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     6661 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     6721 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     6781 gcacatttcc ccgaaaagtg ccacctgacg tc
//
