LOCUS       V039959                 8755 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V039959
VERSION     V039959
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..4245
                     /label="AP3B1(NM_003664)"
                     /note="AP3B1(NM_003664)"
                     /gene="AP3B1"
     CDS             4252..4317
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    4355..4579
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4625..5053
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5067..5396
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5463..6257
                     /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    6431..6552
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6601..6617)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6625..6641
                     /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(6649..6679)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6694..6715
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7003..7588)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7759..8619)
                     /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(8620..8724)
                     /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 accatgtcca gcaatagttt tccttacaat gagcagtccg gaggagggga ggcgacggag
     1021 ctgggtcagg aggcgacctc aaccatttcc ccctcggggg ccttcggcct ctttagcagc
     1081 gatttgaaga agaatgaaga tctaaagcaa atgttagaga gcaacaaaga ttctgctaaa
     1141 ctggatgcta tgaagcggat tgttgggatg attgcaaaag ggaaaaatgc atctgaactg
     1201 tttcctgctg ttgtgaagaa tgtggccagt aaaaatattg agatcaagaa gttggtatat
     1261 gtttacctgg ttcgatatgc tgaagaacag caggatcttg cactcctgtc cataagcact
     1321 tttcagcgag ctctgaagga cccaaaccaa ctaattcgtg caagcgcttt gagagttctg
     1381 tcaagtatta gagtgccaat tattgtacct atcatgatgc ttgctattaa ggaagcttct
     1441 gctgacttat caccatatgt taggaagaat gcagcccatg caatacaaaa attatacagc
     1501 cttgatccag agcagaagga aatgttaatt gaagtaattg aaaaacttct gaaagataaa
     1561 agcacattgg tagctggcag tgttgtgatg gcttttgaag aagtatgccc ggacagaata
     1621 gatctgattc ataaaaatta ccgcaagcta tgtaacttac tagtggatgt tgaagagtgg
     1681 gggcaggttg tcataatcca catgctaact cgatatgctc ggacacagtt tgtcagccct
     1741 tggaaagagg gtgatgaatt agaagacaat ggaaagaatt tctacgaatc tgatgatgat
     1801 cagaaggaaa agactgacaa aaagaagaag ccgtatacta tggatccaga tcatagactc
     1861 ttaattagaa atacaaagcc tttgcttcag agcaggaatg ctgcggtggt tatggcagtt
     1921 gctcagctgt attggcacat atcaccaaaa tctgaagctg gcataatttc taaatcacta
     1981 gtgcgtttac ttcgtagcaa tagggaggtg cagtatattg tcctacaaaa tatagcaact
     2041 atgtcaattc aaagaaaggg gatgtttgaa ccttatctga agagtttcta tgttaggtca
     2101 actgatccaa ctatgatcaa gacactgaag cttgaaattt tgacaaactt ggcaaatgaa
     2161 gccaacatat caactcttct tcgagaattt cagacctatg tgaaaagcca ggataaacaa
     2221 tttgcagcag ccactattca gactataggc agatgtgcaa ccaacatctt ggaagtcact
     2281 gacacgtgcc tcaatggctt ggtctgtctg ctgtccaaca gggatgaaat agttgttgct
     2341 gaaagtgtgg ttgttataaa gaaattactg caaatgcaac ctgcacaaca tggtgaaatt
     2401 attaaacata tggccaaact cctggacagt atcactgttc ctgttgctag agcaagtatt
     2461 ctttggctaa ttggagaaaa ctgtgaacga gttcctaaaa ttgcccctga tgttttgagg
     2521 aagatggcta aaagcttcac tagtgaagat gatctggtaa aactgcagat attaaatctg
     2581 ggagcaaaat tgtatttaac caactccaaa cagacaaaat tgcttaccca gtacatatta
     2641 aatctcggca agtatgatca aaactacgac atcagagacc gtacaagatt tattaggcag
     2701 cttattgttc cgaatgtaaa gagtggagct ttaagtaaat atgccaaaaa aatattccta
     2761 gcacaaaagc ctgcaccact gcttgagtct ccttttaaag atagagatca tttccagctt
     2821 ggcaccttat ctcatactct caacattaaa gctactgggt acctggaatt atctaattgg
     2881 ccagaggtgg cgcccgaccc atcagttcga aatgtagaag taatagagtt ggcaaaagaa
     2941 tggaccccag caggaaaagc aaagcaagag aattctgcta agaagtttta ttctgaatct
     3001 gaggaagagg aggactcttc tgatagtagc agtgacagtg agagtgaatc tggaagtgaa
     3061 agtggagaac aaggcgaaag tggggaggaa ggagacagca atgaggacag cagtgaggac
     3121 tcctccagtg agcaggacag tgagagtgga cgggagtcag gcctagaaaa caaaagaaca
     3181 gccaagagga actcaaaagc caaaggaaaa agtgattctg aagatgggga gaaggaaaat
     3241 gaaaaatcta aaacttcaga ttcttcaaat gacgaatcta gttcaataga agacagttct
     3301 tccgattctg aatcagagtc agaacctgaa agtgaatctg aatccagaag agtcactaag
     3361 gagaaagaaa agaaaacaaa gcaagataga actcctctta ccaaagatgt ttcacttcta
     3421 gatctggatg attttaaccc agtatccact ccagttgcac ttcccacacc agctctttct
     3481 ccaagtttga tggctgatct tgaaggttta cacttgtcaa cttcctcttc agtcatcagt
     3541 gtcagtactc ctgcatttgt accaacgaaa actcacgtgc tgcttcatcg aatgagtgga
     3601 aaaggactag ctgcccatta tttctttcca agacagcctt gcatttttgg tgataagatg
     3661 gtctctatac aaataacact gaataacact actgatcgaa agatagaaaa tatccacata
     3721 ggggaaaaaa aacttcctat aggcatgaaa atgcatgttt ttaatccaat agactctctt
     3781 gagcctgagg gatccattac agtttcaatg ggtattgact tttgtgattc tactcagact
     3841 gccagtttcc agttgtgtac caaggatgat tgcttcaatg ttaatattca gccacctgtt
     3901 ggagaactgc ttttacctgt ggccatgtca gagaaagatt ttaagaaaga gcaaggagtg
     3961 ctaacaggaa tgaatgaaac ttctgctgta atcattgctg caccacagaa tttcactccc
     4021 tctgtgatct ttcagaaggt tgtaaatgta gccaatgtag gtgcagtccc ttctggccag
     4081 gataatatac acaggtttgc agctaaaact gtgcacagtg ggtcattgat gctagtcaca
     4141 gtggaactga aggaaggctc tacagcccag cttatcataa acactgagaa aactgtgatt
     4201 ggctctgttc tgctgcggga actgaagcct gtcctgtctc aggggctcga ggactacaaa
     4261 gaccatgacg gtgattataa agatcatgac atcgactaca aggatgacga tgacaagtag
     4321 tgagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     4381 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     4441 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     4501 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     4561 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     4621 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     4681 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     4741 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     4801 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     4861 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     4921 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     4981 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     5041 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     5101 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     5161 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     5221 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     5281 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     5341 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     5401 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc
     5461 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     5521 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     5581 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     5641 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     5701 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     5761 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     5821 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     5881 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     5941 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     6001 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     6061 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     6121 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     6181 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     6241 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     6301 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     6361 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     6421 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     6481 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     6541 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     6601 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     6661 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     6721 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     6781 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     6841 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     6901 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     6961 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     7021 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     7081 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     7141 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     7201 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     7261 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     7321 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     7381 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     7441 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     7501 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ttttttgttt gcaagcagca
     7561 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     7621 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     7681 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     7741 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     7801 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     7861 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     7921 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     7981 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     8041 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     8101 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     8161 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     8221 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     8281 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     8341 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     8401 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     8461 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     8521 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     8581 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     8641 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     8701 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
