LOCUS       V019131                 8828 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019131
VERSION     V019131
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        431..708
                     /label="chicken β-actin promoter"
     intron          709..1726
                     /note="chimera between introns from chicken β-actin and
                     rabbit β-globin"
                     /label="chimeric intron"
     CDS             1823..3499
                     /label="PRKAA1(NM_006251)"
                     /note="PRKAA1(NM_006251)"
                     /gene="PRKAA1"
     CDS             3515..3580
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3604..4192
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4258..4306
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4508..4936
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4950..5279
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5346..6140
                     /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    6316..6437
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6486..6502)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6510..6526
                     /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(6534..6564)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6579..6600
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6888..7476)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7647..8507)
                     /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(8508..8612)
                     /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 tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc
      481 ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat gggggcgggg
      541 gggggggggg ggcccccccc aggcggggcg gggcggggcg aggggcgggg cggggcgagg
      601 cggaaaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg
      661 aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg agtcgttgcg
      721 cgctgccttc cccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga
      781 ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc gggctgtaat
      841 tagcgcttgg tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag ccttgagggg
      901 ctccgggagg gccctttgtg cggggggagc ggctcggggg gtgcgtgcgt gtgtgtgtgc
      961 gtggggagcg ccgcgtgcgg ctccgcgctg cccggcggct gtgagcgctg cgggcgcggc
     1021 gcggggcttt gtgcgctccg cagtgtgcgc gaggggagcg cggccggggg cggtgccccg
     1081 cggtgcgggg ggggctgcga ggggaacaaa ggctgcgtgc ggggtgtgtg cgtggggggg
     1141 tgagcagggg gtgtgggcgc gtcggtcggg ctgcaacccc ccctgcaccc ccctccccga
     1201 gttgctgagc acggcccggc ttcgggtgcg gggctccgta cggggcgtgg cgcggggctc
     1261 gccgtgccgg gcggggggtg gcggcaggtg ggggtgccgg gcggggcggg gccgcctcgg
     1321 gccggggagg gctcggggga aggggcgcgg cggcccccgg agcgccggcg gctgtcgagg
     1381 cgcggcgagc cgcagccatt gccttttatg gtaatcgtgc gagagggcgc agggacttcc
     1441 tttgtcccaa atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc cctctagcgg
     1501 gcgcggggcg aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg gccttcgtgc
     1561 gtcgccgcgc cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc ggggggacgg
     1621 ctgccttcgg gggggacggg gcagggcggg gttcggcttc tggcgtgtga ccggcggctc
     1681 tagagcctct gctaaccatg ttcatgcctt cttctttttc ctacagctcc tgggcaacgt
     1741 gctggttatt gtgctgtctc atcattttgg caaagaattc tgcagctggc tagcgtttaa
     1801 acttaagctt ggtaccgcca ccatgcgcag actcagttcc tggagaaaga tggcgacagc
     1861 cgagaagcag aaacacgacg ggcgggtgaa gatcggccac tacattctgg gtgacacgct
     1921 gggggtcggc accttcggca aagtgaaggt tggcaaacat gaattgactg ggcataaagt
     1981 agctgtgaag atactcaatc gacagaagat tcggagcctt gatgtggtag gaaaaatccg
     2041 cagagaaatt cagaacctca agcttttcag gcatcctcat ataattaaac tgtaccaggt
     2101 catcagtaca ccatctgata ttttcatggt gatggaatat gtctcaggag gagagctatt
     2161 tgattatatc tgtaagaatg gaaggctgga tgaaaaagaa agtcggcgtc tgttccaaca
     2221 gatcctttct ggtgtggatt attgtcacag gcatatggtg gtccatagag atttgaaacc
     2281 tgaaaatgtc ctgcttgatg cacacatgaa tgcaaagata gctgattttg gtctttcaaa
     2341 catgatgtca gatggtgaat ttttaagaac aagttgtggc tcacccaact atgctgcacc
     2401 agaagtaatt tcaggaagat tgtatgcagg cccagaggta gatatatgga gcagtggggt
     2461 tattctctat gctttattat gtggaaccct tccatttgat gatgaccatg tgccaactct
     2521 ttttaagaag atatgtgatg ggatcttcta tacccctcaa tatttaaatc cttctgtgat
     2581 tagccttttg aaacatatgc tgcaggtgga tcccatgaag agggccacaa tcaaagatat
     2641 cagggaacat gaatggttta aacaggacct tccaaaatat ctctttcctg aggatccatc
     2701 atatagttca accatgattg atgatgaagc cttaaaagaa gtatgtgaaa agtttgagtg
     2761 ctcagaagag gaagttctca gctgtcttta caacagaaat caccaggatc ctttggcagt
     2821 tgcctaccat ctcataatag ataacaggag aataatgaat gaagccaaag atttctattt
     2881 ggcgacaagc ccacctgatt cttttcttga tgatcatcac ctgactcggc cccatcctga
     2941 aagagtacca ttcttggttg ctgaaacacc aagggcacgc catacccttg atgaattaaa
     3001 tccacagaaa tccaaacacc aaggtgtaag gaaagcaaaa tggcatttag gaattagaag
     3061 tcaaagtcga ccaaatgata ttatggcaga agtatgtaga gcaatcaaac aattggatta
     3121 tgaatggaag gttgtaaacc catattattt gcgtgtacga aggaagaatc ctgtgacaag
     3181 cacttactcc aaaatgagtc tacagttata ccaagtggat agtagaactt atctactgga
     3241 tttccgtagt attgatgatg aaattacaga agccaaatca gggactgcta ctccacagag
     3301 atcgggatca gttagcaact atcgatcttg ccaaaggagt gattcagatg ctgaggctca
     3361 aggaaaatcc tcagaagttt ctcttacctc atctgtgacc tcacttgact cttctcctgt
     3421 tgacctaact ccaagacctg gaagtcacac aatagaattt tttgagatgt gtgcaaatct
     3481 aattaaaatt cttgcacaaa tgcggccgct cgaggactac aaagaccatg acggtgatta
     3541 taaagatcat gacatcgact acaaggatga cgatgacaag tagtgagggc ccgatatctc
     3601 gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     3661 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     3721 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     3781 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     3841 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     3901 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     3961 ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg
     4021 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     4081 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     4141 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggaaac
     4201 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     4261 caataaaaag acagaataaa acgcacgggt gttgggtcgt ttgttcataa acgcggggtt
     4321 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
     4381 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
     4441 gccaacgtcg gggcggcagg ccctgccata gcagatctgc gcagctgggg ctctaggggg
     4501 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     4561 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     4621 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc ggggcatccc tttagggttc
     4681 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     4741 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     4801 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     4861 gatttataag ggattttggg gatttcggcc tattggttaa aaaatgagct gatttaacaa
     4921 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     4981 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     5041 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     5101 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     5161 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     5221 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     5281 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt
     5341 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     5401 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     5461 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     5521 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     5581 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     5641 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     5701 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     5761 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     5821 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     5881 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     5941 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     6001 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     6061 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     6121 ttcttgacga gttcttctga gcgggactct ggggttcgcg aaatgaccga ccaagcgacg
     6181 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     6241 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     6301 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     6361 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     6421 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     6481 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     6541 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     6601 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     6661 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     6721 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     6781 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     6841 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     6901 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     6961 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     7021 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     7081 caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     7141 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     7201 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     7261 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     7321 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     7381 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     7441 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     7501 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     7561 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     7621 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     7681 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     7741 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     7801 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     7861 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     7921 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     7981 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     8041 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     8101 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     8161 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     8221 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     8281 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     8341 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     8401 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     8461 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     8521 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     8581 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     8641 gtcgacggat cgggagatct cccgatcccc tatggtcgac tctcagtaca atctgctctg
     8701 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
     8761 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
     8821 tgcttagg
//
