LOCUS       V044261                 7381 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044261
VERSION     V044261
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"
     regulatory      913..922
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             922..987
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             991..2898
                     /label="Pabpc1(NM_008774)"
                     /note="Pabpc1(NM_008774)"
                     /gene="Pabpc1"
     polyA_signal    2978..3202
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3248..3676
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3690..4019
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4086..4880
                     /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    5054..5175
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5224..5240)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5248..5264
                     /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(5272..5302)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5317..5338
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5626..6214)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6385..7245)
                     /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(7246..7350)
                     /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 aagccaccat ggactacaaa gaccatgacg gtgattataa agatcatgac
      961 atcgactaca aggatgacga tgacaagctt atgaacccca gcgcccccag ctaccccatg
     1021 gcctctctgt acgtggggga cctgcacccc gacgtgaccg aggcgatgct ctacgagaag
     1081 ttcagcccgg ccgggcccat cctctccatc cgggtctgca gggacatgat cacccgccgc
     1141 tccttgggct acgcgtacgt gaacttccag cagccggcgg acgcggaacg tgctttggac
     1201 accatgaatt ttgatgttat aaagggcaag ccagtacgca tcatgtggtc tcagcgtgat
     1261 ccatcacttc gcaaaagtgg agtaggcaac atattcatta aaaatttgga caaatccatc
     1321 gacaataaag cactatatga tacgttttct gcgtttggta acatcctttc atgtaaggtg
     1381 gtttgtgatg aaaatggctc caagggctat ggatttgtac actttgaaac acaggaagca
     1441 gctgaaagag ctattgaaaa aatgaatggg atgcttctaa atgatcgtaa agtgtttgtt
     1501 ggacgattta aatctcggaa ggaacgagaa gcagagcttg gagccagggc aaaggagttc
     1561 accaatgttt acatcaagaa ctttggagaa gacatggatg atgagcgcct taaggaactc
     1621 tttggcaagt ttgggcctgc cttaagtgtg aaagtaatga cagatgaaag tggaaaatcc
     1681 aaaggatttg gatttgtaag ctttgaaagg catgaagatg cgcagaaagc tgtggatgag
     1741 atgaatggga aggagctcaa tggaaaacag atttatgttg gacgagctca gaaaaaagtg
     1801 gaacggcaga cggaacttaa gcgcaaattt gagcagatga agcaagatag gatcaccaga
     1861 tatcagggtg tgaaccttta tgtgaaaaat cttgatgacg ggattgatga tgagcgtctc
     1921 cggaaggagt tttctccgtt tggtacaatc accagtgcaa aagtaatgat ggagggtggg
     1981 cgcagcaaag ggtttggttt tgtatgtttc tcatcccctg aagaagccac taaagcagtt
     2041 acagagatga atggtagaat tgtggccacg aagccactgt atgtagcttt agctcagcgc
     2101 aaagaagagc gccaggctca cctcactaac cagtatatgc agaggatggc aagtgtacga
     2161 gctgtgccca accccgtgat caacccctac cagccagcac ctccttcagg ttacttcatg
     2221 gcagctatcc cacagactca gaaccgtgct gcatactatc ctcctagcca aattgctcaa
     2281 ctaagaccaa gtcctcgctg gactgctcag ggtgccagac ctcatccatt ccagaatatg
     2341 cccggtgcta tccgcccagc tgctcctaga ccaccattta gtacgatgag accagcttcc
     2401 tcacaggttc cacgagtcat gtcaacacag cgtgttgcta acacatcaac acagacaatg
     2461 ggtccacgtc ctgcagctgc tgctgctgca gccactcctg ctgtccgcac cgttccccag
     2521 tataaatatg ctgcgggagt ccgcaatccc cagcaacatc ttaatgcaca gccacaagtt
     2581 accatgcaac agcctgctgt tcatgtgcaa ggtcaagaac ctttaactgc ttccatgttg
     2641 gcatctgcgc ccccgcaaga gcagaagcaa atgttgggtg aacggctgtt tcctcttatc
     2701 caagccatgc acccttctct tgctggtaaa atcactggca tgctgttgga gattgataac
     2761 tcagaattac ttcacatgct cgagtctcca gagtctctcc gctcaaaggt tgatgaagct
     2821 gtagctgtac tacaagccca ccaagcgaaa gaggctgccc agaaagcagt gaacagtgcc
     2881 actggtgttc caactgtcta agaattctgc agatatccag cacagtggcg gccgctcgag
     2941 tctagagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
     3001 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     3061 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     3121 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     3181 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     3241 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     3301 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     3361 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     3421 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     3481 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     3541 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     3601 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     3661 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     3721 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     3781 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3841 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     3901 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     3961 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     4021 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt
     4081 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     4141 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     4201 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     4261 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     4321 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     4381 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     4441 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     4501 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     4561 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     4621 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     4681 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     4741 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     4801 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     4861 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     4921 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     4981 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     5041 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     5101 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     5161 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     5221 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     5281 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     5341 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     5401 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     5461 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     5521 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     5581 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5641 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5701 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5761 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5821 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5881 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5941 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     6001 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     6061 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     6121 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     6181 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     6241 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     6301 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     6361 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     6421 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     6481 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     6541 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     6601 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     6661 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     6721 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     6781 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     6841 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     6901 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     6961 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     7021 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     7081 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     7141 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     7201 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     7261 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     7321 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     7381 c
//
