LOCUS       V024461                 6584 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024461
VERSION     V024461
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"
     CDS             941..2074
                     /label="TIMD4(NM_138379)"
                     /note="TIMD4(NM_138379)"
                     /gene="TIMD4"
     CDS             2081..2146
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2184..2408
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2454..2882
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2896..3225
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3292..4086
                     /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    4260..4381
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4430..4446)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4454..4470
                     /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(4478..4508)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4523..4544
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4832..5417)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5588..6448)
                     /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(6449..6553)
                     /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 aagcttggta ccgagctcgg atccgccacc atgtccaaag aacctctcat
      961 tctctggctg atgattgagt tttggtggct ttacctgaca ccagtcactt cagagactgt
     1021 tgtgacggag gttttgggtc accgggtgac tttgccctgt ctgtactcat cctggtctca
     1081 caacagcaac agcatgtgct gggggaaaga ccagtgcccc tactccggtt gcaaggaggc
     1141 gctcatccgc actgatggaa tgagggtgac ctcaagaaag tcagcaaaat atagacttca
     1201 ggggactatc ccgagaggtg atgtctcctt gaccatctta aaccccagtg aaagtgacag
     1261 cggtgtgtac tgctgccgca tagaagtgcc tggctggttc aacgatgtaa agataaacgt
     1321 gcgcctgaat ctacagagag cctcaacaac cacgcacaga acagcaacca ccaccacacg
     1381 cagaacaaca acaacaagcc ccaccaccac ccgacaaatg acaacaaccc cagctgcact
     1441 tccaacaaca gtcgtgacca cacccgatct cacaaccgga acaccactcc agatgacaac
     1501 cattgccgtc ttcacaacag caaacacgtg cctttcacta accccaagca cccttccgga
     1561 ggaagccaca ggtcttctga ctcccgagcc ttctaaggaa gggcccatcc tcactgcaga
     1621 atcagaaact gtcctcccca gtgattcctg gagtagtgtt gagtctactt ctgctgacac
     1681 tgtcctgctg acatccaaag agtccaaagt ttgggatctc ccatcaacat cccacgtgtc
     1741 aatgtggaaa acgagtgatt ctgtgtcttc tcctcagcct ggagcatctg atacagcagt
     1801 tcctgagcag aacaaaacaa caaaaacagg acagatggat ggaataccca tgtcaatgaa
     1861 gaatgaaatg cccatctccc aactactgat gatcatcgcc ccctccttgg gatttgtgct
     1921 cttcgcattg tttgtggcgt ttctcctgag agggaaactc atggaaacct attgttcgca
     1981 gaaacacaca aggctagact acattggaga tagtaaaaat gtcctcaatg acgtgcagca
     2041 tggaagggaa gacgaagacg gcctttttac cctcctcgag gactacaaag accatgacgg
     2101 tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagt gagggcccgt
     2161 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     2221 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     2281 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     2341 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     2401 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc
     2461 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     2521 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     2581 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     2641 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     2701 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     2761 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     2821 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     2881 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     2941 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     3001 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     3061 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3121 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     3181 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     3241 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     3301 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     3361 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     3421 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     3481 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     3541 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     3601 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     3661 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     3721 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     3781 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     3841 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     3901 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     3961 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     4021 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     4081 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     4141 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     4201 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     4261 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     4321 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     4381 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     4441 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     4501 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     4561 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     4621 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     4681 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     4741 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     4801 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     4861 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     4921 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     4981 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     5041 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     5101 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     5161 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     5221 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     5281 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     5341 ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca
     5401 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5461 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5521 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     5581 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     5641 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     5701 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     5761 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     5821 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     5881 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     5941 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     6001 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6061 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6121 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     6181 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     6241 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6301 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6361 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6421 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6481 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6541 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc
//
