LOCUS       V031889                 8652 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031889
VERSION     V031889
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..3332
                     /label="DDX24(NM_020414)"
                     /note="DDX24(NM_020414)"
                     /gene="DDX24"
     CDS             3339..3404
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3428..4016
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4082..4130
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4332..4760
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4774..5103
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5170..5964
                     /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    6140..6261
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6310..6326)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6334..6350
                     /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(6358..6388)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6403..6424
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6712..7300)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7471..8331)
                     /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(8332..8436)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatgaa gttgaaggac acaaaatcaa
      781 ggccaaagca gtcaagctgt ggcaaatttc agacaaaggg aatcaaagtt gtgggaaaat
      841 ggaaggaagt gaagattgac ccaaatatgt ttgcagatgg acagatggat gacttggtgt
      901 gctttgagga attgacagat taccagttgg tctcccctgc caagaatccc tccagtctct
      961 tctcaaagga agcacccaag agaaaggcac aagctgtttc agaagaagag gaggaggagg
     1021 agggaaagtc tagctcacca aagaaaaaga tcaagttgaa gaaaagtaaa aatgtagcaa
     1081 ctgaaggaac cagtacccag aaagaatttg aagtgaaaga tcctgagctg gaggcccagg
     1141 gagatgacat ggtttgtgat gatccggagg ctggggagat gacatcagaa aacctggtcc
     1201 aaactgctcc aaaaaagaag aaaaataaag ggaaaaaagg gttggagcct tctcagagca
     1261 ctgctgccaa ggtgcccaaa aaagcgaaga catggattcc tgaagttcat gatcagaaag
     1321 cagatgtgtc agcttggaag gacctgtttg ttcccaggcc ggttctccga gcactcagct
     1381 ttctaggctt ctctgcaccc acaccaatcc aagccctgac cttggcacct gccatccgtg
     1441 acaaactgga catccttggg gctgctgaga caggaagtgg gaaaactctt gcctttgcca
     1501 tcccaatgat tcatgcggtg ttgcagtggc agaagaggaa tgctgcccct cctccaagta
     1561 acaccgaagc accacctgga gagaccagaa ctgaggccgg agctgagact agatcaccag
     1621 gcaaggctga agctgagtct gatgcattgc ctgacgatac tgtaattgag agtgaagcac
     1681 tgcccagtga tattgcagcc gaggccagag ccaagactgg aggcactgtc tcagaccagg
     1741 cgttgctctt tggtgacgat gatgctggtg aagggccttc ttccctgatc agggagaaac
     1801 ctgttcccaa acagaatgag aatgaggagg aaaatcttga taaagagcag actggaaatc
     1861 taaaacagga gttggatgac aaaagcgcca cctgtaaggc atatccaaag cgtcctctgc
     1921 ttggactggt tctgactccc actcgagagc tggccgtcca ggtcaaacag cacattgatg
     1981 ctgtggccag gtttacagga attaaaactg ctattttggt tggtggaatg tccacgcaga
     2041 aacagcagag gatgctgaac cgtcgtcctg agattgtggt tgctactcca ggccggctgt
     2101 gggaattaat taaagaaaag cattatcatt tgaggaacct tcggcagctc aggtgcctgg
     2161 tagtggatga ggctgaccgg atggttgaga aaggccattt tgctgagctc tcacagctgc
     2221 tagagatgct caatgactcc caatacaacc caaagagaca aacgcttgtt ttttctgcca
     2281 cactcaccct ggtgcatcag gctcctgctc gaatccttca taagaagcac accaagaaaa
     2341 tggataaaac agccaaactt gacctcctta tgcagaaaat tggcatgagg ggcaagccca
     2401 aggtcattga cctcacaagg aatgaggcca cggtggagac gctaacagag accaagatcc
     2461 attgtgagac tgatgagaaa gacttctact tgtactactt cctgatgcag tatccaggcc
     2521 gcagcttagt gtttgccaac agtatctcct gcatcaaacg cctctctggg ctcctcaaag
     2581 tccttgatat catgcccttg accctgcatg cctgtatgca ccagaagcag aggctcagaa
     2641 acctggagca gtttgcccgt ctggaagact gtgttctctt ggcaacagat gtggcagctc
     2701 ggggtctgga tattcctaaa gtccagcatg tcatccatta ccaggtccca cgtacctcgg
     2761 agatttatgt ccaccgaagt ggtcgaactg ctcgagctac caatgaaggc ctcagtctga
     2821 tgctcattgg gcctgaggat gtgatcaact ttaagaagat ttacaaaacg ctcaagaaag
     2881 atgaggatat cccactgttc cccgtgcaga caaaatacat ggatgtggtc aaggagcgaa
     2941 tccgtttagc tcgacagatt gagaaatctg agtatcggaa cttccaggct tgcctgcaca
     3001 actcttggat tgagcaggca gcagctgccc tggagattga gctggaagaa gacatgtata
     3061 agggaggaaa agctgaccag caagaagaac gtcggagaca aaagcagatg aaggttctga
     3121 agaaggagct gcgccacctg ctgtcccagc cactgtttac ggagagccag aaaaccaagt
     3181 atcccactca gtctggcaag ccgcccctgc ttgtgtctgc cccaagtaag agcgagtctg
     3241 ctttgagctg tctctccaag cagaagaaga agaagacaaa gaagccgaag gagccacagc
     3301 cggaacagcc acagccaagt acaagtgcaa atggatccga ctacaaagac catgacggtg
     3361 attataaaga tcatgacatc gactacaagg atgacgatga caagtagtga gggcccgata
     3421 tctcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     3481 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     3541 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     3601 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     3661 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     3721 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     3781 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg
     3841 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     3901 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     3961 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg
     4021 aaacggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     4081 acggcaataa aaagacagaa taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg
     4141 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     4201 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     4261 cgcagccaac gtcggggcgg caggccctgc catagcagat ctgcgcagct ggggctctag
     4321 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     4381 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     4441 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcggggca tccctttagg
     4501 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     4561 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     4621 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     4681 ttttgattta taagggattt tggggatttc ggcctattgg ttaaaaaatg agctgattta
     4741 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     4801 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     4861 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4921 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     4981 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     5041 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     5101 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     5161 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     5221 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     5281 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     5341 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     5401 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     5461 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     5521 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     5581 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     5641 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     5701 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     5761 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     5821 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     5881 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     5941 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgcgaaatga ccgaccaagc
     6001 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     6061 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     6121 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     6181 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     6241 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     6301 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     6361 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     6421 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     6481 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     6541 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     6601 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     6661 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     6721 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     6781 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     6841 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     6901 ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     6961 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     7021 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     7081 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     7141 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     7201 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     7261 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     7321 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     7381 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     7441 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     7501 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     7561 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     7621 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     7681 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     7741 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     7801 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     7861 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     7921 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     7981 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     8041 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     8101 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     8161 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     8221 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     8281 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     8341 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     8401 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     8461 tgacgtcgac ggatcgggag atctcccgat cccctatggt cgactctcag tacaatctgc
     8521 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
     8581 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
     8641 aatctgctta gg
//
