LOCUS       V019124                 6739 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019124
VERSION     V019124
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             928..1017
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     CDS             1039..2292
                     /label="PELI1(NM_020651)"
                     /note="PELI1(NM_020651)"
                     /gene="PELI1"
     polyA_signal    2339..2563
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2609..3037
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3051..3380
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3447..4241
                     /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    4415..4536
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4585..4601)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4609..4625
                     /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(4633..4663)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4678..4699
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4987..5572)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5743..6603)
                     /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(6604..6708)
                     /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 ggcggcggaa caaaaactaa tatcagagga agatttggaa
      961 cagaaactaa tcagtgaaga agatttagag caaaagttaa tttccgagga ggacttatca
     1021 gcggaaaagc ttggtaccat gttttctcct gatcaagaaa atcatccatc taaagcacca
     1081 gtaaaatatg gtgaactcat tgtcttaggg tataatgggt ctctcccaaa tggcgataga
     1141 ggaaggagga aaagtaggtt tgctttgttt aaaagaccta aggcaaatgg ggtgaagccc
     1201 agcactgtgc atattgcttg tactcctcag gctgcaaagg caataagcaa caaagaccag
     1261 catagcatat catatacttt atctcgggcc cagactgtgg tggttgaata tactcatgac
     1321 agcaacaccg atatgtttca gattggccgg tcgactgaaa gccccattga ttttgtagta
     1381 actgacacgg ttcctggaag tcaaagtaat tctgatacac agtcagtaca aagcactata
     1441 tcaagatttg cctgcagaat catatgtgaa cggaatcctc cctttacagc acggatttat
     1501 gctgcaggat ttgactcatc aaaaaacatc tttcttgggg agaaggctgc caaatggaag
     1561 cgatcagatg gacagatgga tggcttgacc actaatggtg ttcttgtgat gcatccacgc
     1621 aatgggttca cagaagactc caagcctgga atatggagag aaatatcggt gtgtggaaat
     1681 gtatttagcc tacgtgaaac cagatcggct cagcagagag gaaaaatggt ggaaattgaa
     1741 accaatcagt tacaagatgg ctcgttaatt gacctctgtg gtgcaacatt gttatggcgt
     1801 actgcagaag gcctttccca cactcctcgc gtgaagcatt tagaagcttt aagacaggaa
     1861 atcaatgcag cacgacctca gtgccctgta gggttcaaca cactagcatt tcctagtatg
     1921 aagaggaaag acgttgtaga tgaaaaacaa ccatgggtat atctaaactg cggccatgta
     1981 catggctatc ataactgggg aaacaaagaa gaacgtgatg gaaaagatcg tgaatgtcct
     2041 atgtgtaggt ctgttggtcc ctatgttcct ctgtggcttg gatgtgaagc tggattttat
     2101 gtggacgccg gccctccaac ccatgcgttt agcccgtgtg ggcatgtgtg ttcagaaaag
     2161 acaactgcct attggtccca gatcccactt cctcatggta ctcatacttt tcatgcagcc
     2221 tgtccctttt gtgcacatca gttggctggt gaacaaggct acatcagact tatttttcaa
     2281 ggacctctag actaactcga gtctagaggg cccgtttaaa cccgctgatc agcctcgact
     2341 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     2401 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     2461 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     2521 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     2581 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     2641 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     2701 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     2761 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     2821 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     2881 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     2941 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     3001 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     3061 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3121 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3181 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     3241 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     3301 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     3361 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     3421 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     3481 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     3541 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     3601 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     3661 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     3721 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     3781 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     3841 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     3901 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     3961 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     4021 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     4081 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     4141 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     4201 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     4261 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     4321 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     4381 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     4441 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     4501 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     4561 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     4621 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     4681 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     4741 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     4801 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     4861 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     4921 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     4981 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     5041 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     5101 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     5161 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     5221 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     5281 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     5341 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     5401 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     5461 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     5521 cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     5581 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     5641 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     5701 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     5761 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     5821 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     5881 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     5941 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     6001 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     6061 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     6121 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     6181 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     6241 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     6301 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     6361 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     6421 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     6481 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     6541 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     6601 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     6661 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     6721 aaaagtgcca cctgacgtc
//
