LOCUS       V041814                 7252 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041814
VERSION     V041814
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             964..2742
                     /label="ARSD(NM_001669)"
                     /note="ARSD(NM_001669)"
                     /gene="ARSD"
     CDS             2749..2814
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2852..3076
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3122..3550
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3564..3893
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3960..4754
                     /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    4928..5049
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5098..5114)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5122..5138
                     /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(5146..5176)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5191..5212
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5500..6085)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6256..7116)
                     /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(7117..7221)
                     /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 atccactagt ccagtgtggt ggaattcgcc
      961 accatgcgat ccgccgcgcg gaggggacgc gccgcgcccg ccgccaggga ctctttgccg
     1021 gtgctactgt ttttatgctt gcttctgaag acgtgtgaac ctaaaactgc aaatgccttt
     1081 aaaccaaata tcctactgat catggcggat gatctaggca ctggggatct cggttgctac
     1141 gggaacaata cactgagaac gccgaatatt gaccagcttg cagaggaagg tgtgaggctc
     1201 actcagcacc tggcggccgc cccgctctgc accccaagcc gagctgcatt cctcacaggg
     1261 agacattcct tcagatcagg catggacgcc agcaatggat accgggccct tcagtggaac
     1321 gcaggctcag gtggactccc tgagaacgaa accacttttg caagaatctt gcagcagcat
     1381 ggctatgcaa ccggcctcat aggaaaatgg caccagggtg tgaattgtgc atcccgcggg
     1441 gatcactgcc accaccccct gaaccacgga tttgactatt tctacggcat gcccttcacg
     1501 ctcacaaacg actgtgaccc aggcaggccc cccgaagtgg acgccgccct gagggcgcag
     1561 ctctggggtt acacccagtt cctggcgctg gggattctca ccctggctgc cggccagacc
     1621 tgcggtttct tctctgtctc cgcgagagca gtcaccggca tggccggcgt gggctgcctg
     1681 tttttcatct cttggtactc ctccttcggg tttgtgcgac gctggaactg tatcctgatg
     1741 agaaaccatg acgtcacgga gcaacccatg gttctggaga aaacagcgag tcttatgcta
     1801 aaggaagctg tttcctatat tgaaagacac aagcatgggc catttctcct cttcctttct
     1861 ttgctgcatg tgcacattcc ccttgtgacc acgagtgcat tcctggggaa aagtcagcat
     1921 ggcttatatg gtgataatgt ggaggagatg gactggctca taggtaaggt tcttaatgcc
     1981 atcgaagaca atggtttaaa gaactcaaca ttcacgtatt tcacctctga ccatggagga
     2041 catttagagg caagagatgg acacagccag ttagggggat ggaacggaat ttacaaaggt
     2101 gggaagggca tgggaggatg ggaaggtggg atccgcgtgc ccgggatctt ccactggccg
     2161 ggggtgctcc cggccggccg agtgattgga gagcccacga gcctgatgga cgtgttccct
     2221 actgtggtcc agctggtggg tggcgaggtg ccccaggaca gggtgattga tggccacagc
     2281 ctggtaccct tgctgcaggg agctgaggca cgctcggcac atgagttcct gtttcattac
     2341 tgtgggcagc atcttcacgc agcacgctgg caccagaagg acagtggaag cgtctggaag
     2401 gttcattaca cgaccccgca gttccacccc gagggagcgg gggcctgcta cggccgaggc
     2461 gtctgcccat gctccgggga gggcgtgacc catcacagac cccctttgct ctttgacctc
     2521 tccagggacc cctccgaggc acggcccctg acccccgact ccgagcccct gtaccacgcc
     2581 gtgatagcaa gggtaggtgc cgcggtgtcg gagcatcggc agaccctgag tcctgtgccc
     2641 cagcagtttt ccatgagcaa catcctgtgg aagccgtggc tgcagccgtg ctgcggacat
     2701 ttcccgttct gttcatgcca cgaggatggg gatggcaccc ccctcgagga ctacaaagac
     2761 catgacggtg attataaaga tcatgacatc gactacaagg atgacgatga caagtagtga
     2821 gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     2881 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     2941 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     3001 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     3061 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     3121 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     3181 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     3241 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     3301 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     3361 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     3421 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     3481 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     3541 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     3601 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     3661 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3721 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     3781 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     3841 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     3901 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
     3961 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     4021 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     4081 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     4141 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     4201 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     4261 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     4321 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     4381 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     4441 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     4501 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     4561 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     4621 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     4681 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     4741 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     4801 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     4861 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     4921 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     4981 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     5041 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     5101 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     5161 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     5221 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     5281 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     5341 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     5401 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     5461 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5521 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5581 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5641 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     5701 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5761 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5821 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5881 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5941 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     6001 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca agcagcagat
     6061 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     6121 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     6181 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     6241 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     6301 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     6361 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     6421 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     6481 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     6541 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     6601 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     6661 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     6721 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     6781 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     6841 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     6901 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6961 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     7021 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     7081 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     7141 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     7201 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
