LOCUS       V020821                 7519 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020821
VERSION     V020821
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      901..910
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             910..933
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             943..3072
                     /label="Immt(NM_001034928)"
                     /note="Immt(NM_001034928)"
                     /gene="Immt"
     polyA_signal    3119..3343
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3389..3817
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3831..4160
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4227..5021
                     /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    5195..5316
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5365..5381)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5389..5405
                     /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(5413..5443)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5458..5479
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5767..6352)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6523..7383)
                     /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(7384..7488)
                     /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 gccaccatgg attacaagga tgacgacgat aagactaagc ttatgctgcg ggcctgtcag
      961 ttatccggcg tgaccgtcgc ggcccagagt tgtctgtgtg ggaagtttgt cctccgccca
     1021 ctacgaccat gccgtagata ctccacttca agcagttctg gggtgactgc tggtaaaatt
     1081 gctggagctg gccttttgtt tgttggtgga gggattggtg gcactatcct gtatgccaaa
     1141 tgggattccc atttccggga aagtgtagaa aaaactattc cttattcaga caagctcttt
     1201 gggatggttc ttggctctgc accttatact gttccattgc caaagaaacc gattcagtct
     1261 ggtccactaa aaatctccag cgtatcagag gtgatgacag actctgaact gcccatggca
     1321 cagacccagg aaacgaacgg ggatactcca gcttcagcag caggtgatcc ggcccctgag
     1381 gttgagcatg aggacactat aaacaccgaa tgtcccaaca ctgatgaagg aacatctaca
     1441 ttcgtaactg cagctctagc caagagttta gaagatgctc tgaaccagac tgcaaccgtc
     1501 actcggcaga ctattacagc ccagaatgct gcagttcagg ctgtcaaggc acactccagc
     1561 acactgaaaa cagccatgga caattctgag attgcaggtg agaagaagtc agcccagtgg
     1621 cgaacagtgg agggtgcgtt gaaggagcga agaaaggcag ttgacgaagc tgctgatgcc
     1681 cttctcaaag ccaaagaaga gttagagaag atgaaaacta taattgaaga tgcaaagaaa
     1741 agagagattg ctggggccac cccttatata actgctgcag aggagaaact gcacagcatg
     1801 atagtggatc tggatagcgt ggtcaaaaag gtccaggctg ctcagtcaga ggctaaggtg
     1861 gtgtctcaat atcatgaact tgtggtccaa gcaagggatg actttagaaa agagctggac
     1921 agcatcactc cagacatcac tcctggatgg aaagggatga gtatttctga cctagctggc
     1981 acactctcca ctgacgacct gaacgcgctc attgctcatg cccaccgtcg cattgatcag
     2041 ctcaacagag agctagcgca acagaaagcc acagagaagc agcacatcga gctagccctg
     2101 gaaaggcaaa agctagaaga aaagcgagcc tttgactctg cggtggcaaa agcgttggag
     2161 caccatagaa gtgaaattca ggctgagcaa gacagaaagg tagaggaagt cagagatgcc
     2221 atggagaatg aaatgcggac ccaacttcga cgacaggcag ctgcccacac tgatcatttg
     2281 cgagatgtcc tcaaggtgca agaacaggaa ctgaagtttg aatttgaaca ggacctgtct
     2341 gagaaattgt ctgagcaaga attagagttt catcgtcgca gtcaagaaca aatggacaac
     2401 tttactctgg acataaatac tgcctatgca aggttgagag gaattgagca ggctgttcag
     2461 agtcatgcag ttgctgaaga agaagccaga aaagcccacc agctctggct ttcagtggag
     2521 gcattaaagt acagcatgaa gacctcatct gcagaaatgc ctaccatccc actggggagt
     2581 gcagttgagg ccatcagagt cagctgttct gataatgaat ttacccaggc tttaactgca
     2641 gccattcctc cagagtccct gacccggggg gtatacagtg aagagaccct aagagcccgt
     2701 ttctatgctg ttcaaaaact agctggaagg gtagcgatga ttgatgaaac caaaaatagc
     2761 ctgtaccaat acttcctttc ctacttacag tccctacttt tgttcccgcc taagcaactg
     2821 aagcctccgg cagagctcta ccctgaagat ataaatacat ttaaattgct ctcatatgcc
     2881 tcctattgta ttgagcatgg tgacctggag ctggccgcaa agtttgtcaa tcagctaaaa
     2941 ggggagtcca gaagagtggc acaggactgg ctgaaggagg cccgaatgac cttagaaact
     3001 aagcagatag tggagatcct gactacatac gccagcgccg taggaatagg aaccactcaa
     3061 gtgcagcaag agtaactcga gtctagaggg cccgtttaaa cccgctgatc agcctcgact
     3121 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     3181 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     3241 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     3301 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     3361 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     3421 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     3481 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     3541 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     3601 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     3661 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     3721 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     3781 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     3841 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3901 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3961 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     4021 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     4081 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     4141 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     4201 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     4261 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     4321 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     4381 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     4441 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     4501 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     4561 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     4621 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     4681 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     4741 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     4801 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     4861 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     4921 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     4981 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     5041 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     5101 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     5161 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     5221 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     5281 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     5341 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     5401 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     5461 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     5521 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     5581 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     5641 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     5701 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     5761 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     5821 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     5881 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     5941 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6001 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6061 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6121 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     6181 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     6241 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6301 cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     6361 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     6421 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     6481 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     6541 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     6601 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     6661 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     6721 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     6781 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     6841 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     6901 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     6961 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     7021 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     7081 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     7141 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     7201 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     7261 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     7321 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     7381 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     7441 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     7501 aaaagtgcca cctgacgtc
//
