LOCUS       V021862                 7009 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021862
VERSION     V021862
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             1009..2529
                     /label="THUMPD3(NM_001114092)"
                     /note="THUMPD3(NM_001114092)"
                     /gene="THUMPD3"
     polyA_signal    2609..2833
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2879..3307
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3321..3650
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3717..4511
                     /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    4685..4806
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4855..4871)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4879..4895
                     /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(4903..4933)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4948..4969
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5257..5842)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6013..6873)
                     /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(6874..6978)
                     /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 ggtaccgagc tcggatccat gtgtgacatt
     1021 gaagaagcca ctaaccaact cctagatgtg aaccttcatg agaaccagaa gtctgtacaa
     1081 gtgacagaaa gtgacctcgg aagtgaatct gagcttctag tcactattgg agccactgta
     1141 cctactggct ttgagcaaac agctgcagat gaagtcagag agaaacttgg gtcatcatgc
     1201 aaaatcagca gagaccgtgg caagatatat tttgtcattt cagtggaaag tctggcacag
     1261 gttcattgtc tgagatcagt tgataactta tttgtggtgg ttcaggagtt tcaagattac
     1321 cagttcaaac aaacaaagga agaagttcta aaggattttg aagacttggc tggaaaactc
     1381 ccatggtcaa accccttaaa agtgtggaaa attaatgcca gttttaaaaa gaaaaaagca
     1441 aagcgcaaaa agataaatca gaattcaagt aaagagaaga ttaataatgg acaagaagtc
     1501 aaaatcgatc agagaaatgt taaaaaagag ttcactagcc atgctttaga ttctcatatc
     1561 ttagattatt atgaaaatcc agccatcaaa gaggatgtat caacattaat aggtgatgat
     1621 ttggcatctt gcaaagatga gactgatgaa agctcaaaag aagaaactga gcctcaagtg
     1681 ctgaagttta gagtcacatg caacagggca ggagagaaac attgctttac ctcaaatgag
     1741 gctgcaagag attttggggg tgctgttcaa gattatttta agtggaaggc cgacatgacc
     1801 aactttgatg tggaggttct tttgaacatc catgataatg aagtcattgt gggcattgca
     1861 ttgactgaag agagtctcca ccgaagaaat ataacacatt ttggacctac aactcttaga
     1921 tcaactcttg cctatgggat gctcaggctc tgtgatcctc taccttatga tataatagtc
     1981 gatccaatgt gtggaactgg ggcaatacca atagaggggg ccactgaatg gtctgactgt
     2041 ttccatattg ctggtgataa taatccactg gctgtgaata gagcagcaaa taacattgca
     2101 tctttattga ccaagagcca aattaaagaa ggcaaaccct cctggggctt gcccatagat
     2161 gctgttcagt gggatatctg caatctgcca ttgagaactg gctctgtgga tattattgta
     2221 acagatttgc catttggaaa aaggatggga tccaagaaga gaaactggaa cctttatcca
     2281 gcttgcctac gggagatgag ccgtgtctgc acacctacca caggccgagc tgtactactt
     2341 actcaagaca caaaatgctt taccaaggcg ttatctggaa tgcgacacgt atggcgaaag
     2401 gtggatacag tctgggtgaa cgttggtggt cttcgtgctg cagtttacgt tctgatacgt
     2461 acacctcaag cttttgttca tccttcagaa caagacggag aaagaggaac tctttggcaa
     2521 tgcaaagaat gagaattctg cagatatcca gcacagtggc ggccgctcga gtctagaggg
     2581 cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     2641 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     2701 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     2761 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     2821 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     2881 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     2941 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     3001 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     3061 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     3121 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     3181 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     3241 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     3301 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     3361 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     3421 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     3481 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     3541 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     3601 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     3661 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     3721 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     3781 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     3841 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     3901 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     3961 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     4021 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     4081 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     4141 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     4201 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     4261 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     4321 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     4381 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     4441 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     4501 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     4561 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     4621 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     4681 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     4741 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4801 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     4861 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     4921 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     4981 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     5041 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     5101 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     5161 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     5221 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     5281 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     5341 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     5401 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     5461 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     5521 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     5581 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     5641 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     5701 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     5761 ttgatccggc aaacaaacca ccgctggtag cggttttttt gtttgcaagc agcagattac
     5821 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     5881 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     5941 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     6001 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     6061 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     6121 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     6181 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     6241 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     6301 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     6361 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     6421 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     6481 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     6541 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     6601 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     6661 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     6721 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     6781 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     6841 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     6901 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     6961 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
