LOCUS       V038551                 6928 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V038551
VERSION     V038551
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             964..2418
                     /label="TRIM68(NM_018073)"
                     /note="TRIM68(NM_018073)"
                     /gene="TRIM68"
     CDS             2425..2490
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2528..2752
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2798..3226
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3240..3569
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3636..4430
                     /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    4604..4725
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4774..4790)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4798..4814
                     /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(4822..4852)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4867..4888
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5176..5761)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5932..6792)
                     /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(6793..6897)
                     /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 atccactagt ccagtgtggt ggaattcgcc
      961 accatggatc ccacagcctt ggtggaagcc attgtggaag aagtggcctg tcccatctgt
     1021 atgaccttcc tgagggagcc catgagcatt gactgtggcc acagcttctg ccacagctgt
     1081 ctctctggac tctgggagat cccaggagaa tcccagaact ggggttacac ctgtcccctc
     1141 tgtcgagctc ctgtccagcc aaggaacctg cggcctaatt ggcagctggc caatgttgta
     1201 gaaaaagtcc gtctgctaag gctacatcca ggaatggggc tgaagggtga cctgtgtgag
     1261 cgccatgggg aaaagctgaa gatgttctgc aaagaggatg tcttgataat gtgtgaggcc
     1321 tgcagccagt ccccagagca tgaggcccac agtgttgtgc caatggagga tgttgcctgg
     1381 gagtacaagt gggaacttca tgaggccctc gaacatctga agaaagagca agaagaggcc
     1441 tggaagcttg aagttggtga aaggaaacga actgccacct ggaagataca ggtggaaacc
     1501 cgaaaacaga gtattgtatg ggagtttgaa aaataccagc gattactaga gaaaaagcag
     1561 ccaccacatc ggcagctggg ggcagaggta gcagcagctc tggccagcct acagcgggag
     1621 gcagcggaga ccatgcagaa actggagttg aaccatagcg agctcatcca gcagagccag
     1681 gtcctgtgga ggatgattgc agagttgaaa gagaggtcgc agaggcctgt ccgctggatg
     1741 ttgcaggata ttcaggaagt gttaaacagg agcaaatctt ggagcttgca gcagccagaa
     1801 ccaatctccc tggagttgaa gacagattgc cgtgtgctgg ggctaagaga gatcctgaag
     1861 acttatgcag ctgatgtgcg cttggatcca gatactgctt actcccgtct catcgtgtct
     1921 gaggacagaa aacgtgtgca ctatggagac accaaccaga aactgccaga caatcctgag
     1981 agattttacc gctataatat cgtcctggga agccagtgca tctcctcagg ccggcactac
     2041 tgggaggtgg aggtgggaga caggtctgag tggggcctgg gagtatgtaa gcaaaatgta
     2101 gaccggaagg aggtggtcta cttatccccc cactatggat tctgggtgat aaggctgagg
     2161 aagggaaatg agtaccgagc aggcaccgat gagtacccaa tcctgtcctt gccggtccct
     2221 cctcgccggg tgggaatctt cgtggattat gaggcccatg acatttcttt ctacaatgtg
     2281 actgactgtg gctcccacat cttcactttc ccccgctatc ccttccctgg gcgcctcctg
     2341 ccctatttta gtccttgcta cagcattgga accaacaaca ctgctcctct ggccatctgc
     2401 tccctggatg gggaggacct cgaggactac aaagaccatg acggtgatta taaagatcat
     2461 gacatcgact acaaggatga cgatgacaag tagtgagggc ccgtttaaac ccgctgatca
     2521 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     2581 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     2641 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     2701 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     2761 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     2821 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     2881 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     2941 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     3001 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     3061 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     3121 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     3181 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     3241 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3301 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3361 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     3421 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3481 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     3541 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     3601 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     3661 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     3721 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     3781 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     3841 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     3901 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     3961 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     4021 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     4081 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     4141 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     4201 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     4261 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     4321 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     4381 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     4441 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     4501 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     4561 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     4621 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     4681 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     4741 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     4801 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     4861 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     4921 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     4981 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     5041 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     5101 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     5161 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     5221 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     5281 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     5341 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     5401 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     5461 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     5521 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     5581 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     5641 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     5701 cgctggtagc ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     5761 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     5821 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     5881 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     5941 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     6001 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     6061 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     6121 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     6181 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     6241 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     6301 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     6361 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     6421 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     6481 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     6541 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     6601 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     6661 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     6721 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     6781 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     6841 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     6901 atttccccga aaagtgccac ctgacgtc
//
