LOCUS       V029356                 6433 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029356
VERSION     V029356
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             706..1089
                     /label="UBA52(NM_001033930)"
                     /note="UBA52(NM_001033930)"
                     /gene="UBA52"
     CDS             1096..1185
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    1209..1797
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    1863..1911
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2113..2541
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2555..2884
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             2951..3745
                     /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    3921..4042
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4091..4107)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4115..4131
                     /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(4139..4169)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4184..4205
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4493..5081)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5252..6112)
                     /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(6113..6217)
                     /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 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccacta gtccagtgtg gtggaattcg ccaccatgca gatctttgtg
      721 aagaccctca ctggcaaaac catcaccctt gaggtcgagc ccagtgacac cattgagaat
      781 gtcaaagcca aaattcaaga caaggagggt atcccacctg accagcagcg tctgatattt
      841 gccggcaaac agctggagga tggccgcact ctctcagact acaacatcca gaaagagtcc
      901 accctgcacc tggtgttgcg cctgcgaggt ggcattattg agccttctct ccgccagctt
      961 gcccagaaat acaactgcga caagatgatc tgccgcaagt gctatgctcg ccttcaccct
     1021 cgtgctgtca actgccgcaa gaagaagtgt ggtcacacca acaacctgcg tcccaagaag
     1081 aaggtcaaac tcgaggaaca aaaactaata tcagaggaag atttggaaca gaaactaatc
     1141 agtgaagaag atttagagca aaagttaatt tccgaggagg acttatagtg agggcccgat
     1201 atctcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     1261 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     1321 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     1381 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     1441 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
     1501 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     1561 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat
     1621 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     1681 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
     1741 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg
     1801 gaaacggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat
     1861 gacggcaata aaaagacaga ataaaacgca cgggtgttgg gtcgtttgtt cataaacgcg
     1921 gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat
     1981 acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc
     2041 tcgcagccaa cgtcggggcg gcaggccctg ccatagcaga tctgcgcagc tggggctcta
     2101 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     2161 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     2221 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggc atccctttag
     2281 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     2341 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     2401 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     2461 cttttgattt ataagggatt ttggggattt cggcctattg gttaaaaaat gagctgattt
     2521 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
     2581 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     2641 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     2701 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     2761 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     2821 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     2881 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg
     2941 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     3001 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     3061 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     3121 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     3181 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     3241 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     3301 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     3361 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     3421 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     3481 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     3541 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     3601 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     3661 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     3721 tcgccttctt gacgagttct tctgagcggg actctggggt tcgcgaaatg accgaccaag
     3781 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     3841 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     3901 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     3961 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     4021 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     4081 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     4141 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     4201 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     4261 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     4321 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     4381 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     4441 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     4501 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     4561 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     4621 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     4681 tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     4741 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     4801 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     4861 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     4921 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     4981 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     5041 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     5101 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     5161 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     5221 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     5281 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     5341 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     5401 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     5461 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     5521 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     5581 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     5641 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     5701 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     5761 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     5821 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     5881 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     5941 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     6001 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     6061 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     6121 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     6181 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     6241 ctgacgtcga cggatcggga gatctcccga tcccctatgg tcgactctca gtacaatctg
     6301 ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga
     6361 gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa
     6421 gaatctgctt agg
//
