LOCUS       V044323                 8142 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044323
VERSION     V044323
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..2822
                     /label="Fastkd2(NM_172422)"
                     /note="Fastkd2(NM_172422)"
                     /gene="Fastkd2"
     CDS             2829..2894
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2918..3506
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3572..3620
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3822..4250
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4264..4593
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4660..5454
                     /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    5630..5751
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5800..5816)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5824..5840
                     /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(5848..5878)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5893..5914
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6202..6790)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6961..7821)
                     /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(7822..7926)
                     /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 tagcaaagca cgttccttgt
      781 tatggaccat ccgacgattc agtacgttac ttccaagaag cagagctttg aggatagatc
      841 cattgggaac ttgcagacca gaggttattc attcaaaatg gaacccaaga aaccatcggc
      901 taaatgtttt tgatgaggga ttgcaaccat ctgttagata cctctttcag gatatattta
      961 tttctaaatc agtagatggt tgtattcaaa ctaaaggcat aagccattca gccgttttta
     1021 aacctgatag gctgctttgt cctagaagac tgtcctttga tgcaaaacat tcttttgtct
     1081 ctgatgggac atctgatcat gatttaaaga aaataaactt tcatcatacc tccagtgaag
     1141 atgtgttcac caagaaagta agaccaaccc ctgtgaatta taaaaagcta gcccaggagt
     1201 gtaactctct gagtgatgtg ctggacacat tttcaaaagc acctacattt cctggtagta
     1261 actatttctt agcaatgtgg ataattgcca aaaggatatc tgaagacaag aggcgctttg
     1321 agagacagct gatgttcagt caccctgcct ttaaccagct gtgtgagcag atgatgagag
     1381 aagccaagat tatgcactat gaccacctct tgttcagcct taatgctata gtgaagctgg
     1441 gaatccctca gaacactctt atggtacaga ctttgctgag gacgattcag gagcgcatca
     1501 atgagtgtga tgagagatgt ctttcaattt tgtcaaccgc tttagtgtca atggagccat
     1561 gcatgaatgt gaatgcgctt cgagcagggt tgcgaatcct agttgatcag caagtttgga
     1621 acataaaaca tgtcttcacc ttacaaacag tgatgaaatg tattgggaaa gacgcaccat
     1681 ctgcacttaa gaagaaattg gagatgaaag ccttgaagga gttaggcaga ttttccatct
     1741 tgaatagcca gcacatgttt gaagtcttag cagctatgga tctccgctct gttgtccttc
     1801 tgaatgaatg cagtaaggtt gtcatagata atgtccatgg gtgtcctttt aaagtattga
     1861 tcagcatact gcagtcctgt agagacctcc gataccaaaa tgaagatctc ttcaaaagca
     1921 tagcagaata tgtggctaca acctttgaca tctggaagtt gaaacaagtt atctttttcc
     1981 tcttattatt tgaaactctt ggttttcgac ctcctggttt gatggacaag cttatggaga
     2041 aagtagtaca ggagcctggc tctctaaacg tgaagaacat cgtttctatc ctgcatgtgt
     2101 attcttctct caatcatgtt cacaaaatcc acaacagaga gttcctagaa gctctggcga
     2161 gtgctctgac tggctgtctt caccacattt cttctgaaag cctgttgaat gctgtgcatt
     2221 cgttttgcat gatgaattat ttccctctgg cccctattaa ccagcttatc aaagagaata
     2281 tcatcaacga actgctgact tcgggtgaca cagagaagaa tattcataag cttcatgttt
     2341 tgaatacttg tctgaaactt gatgaaagta cttacaaatc tgtgcacata ccgttgccac
     2401 agctgccact gtcagcatca cagccaaacg agaagcttgc agaggtcctc agcaggcttc
     2461 tggaagggga aggacgcttc tcgagaaatg tgccattgcc acataattat cacattgatt
     2521 ttgaaatcag aatggatact aacaggaccc aagtattttc attttctgat gtagatgcaa
     2581 gttctgccac aaacatgcag agagtagcag tgctgtgtgt tcctaagtct gtttactgtt
     2641 taaattcatg ccaccccaga ggattgatgg ccatgaaaat ccgacatttg aatgtaatgg
     2701 gcttccatgt gatcttgatc cataactggg agctgaagaa actaaagatg gaggatgcgg
     2761 tcacatttgt gaggaagaag atctattcag atgaagtgct tcccactgct gacacaaccg
     2821 tgggatccga ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg
     2881 atgacgatga caagtagtga gggcccgata tctcgacaat caacctctgg attacaaaat
     2941 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     3001 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     3061 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     3121 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     3181 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     3241 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     3301 gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct
     3361 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     3421 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     3481 gatctccctt tgggccgcct ccccgcctgg aaacggggga ggctaactga aacacggaag
     3541 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac
     3601 gggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     3661 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     3721 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     3781 catagcagat ctgcgcagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     3841 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     3901 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     3961 tcaagctcta aatcggggca tccctttagg gttccgattt agtgctttac ggcacctcga
     4021 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     4081 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     4141 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tggggatttc
     4201 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     4261 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4321 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     4381 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     4441 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     4501 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     4561 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     4621 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     4681 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     4741 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     4801 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     4861 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     4921 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     4981 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     5041 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     5101 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     5161 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     5221 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     5281 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     5341 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     5401 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     5461 ctctggggtt cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     5521 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     5581 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     5641 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     5701 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     5761 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     5821 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     5881 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     5941 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     6001 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     6061 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     6121 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     6181 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     6241 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     6301 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     6361 cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag
     6421 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     6481 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     6541 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     6601 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     6661 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     6721 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     6781 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     6841 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     6901 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     6961 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     7021 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     7081 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     7141 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     7201 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     7261 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     7321 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     7381 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     7441 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     7501 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     7561 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     7621 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     7681 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     7741 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     7801 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     7861 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     7921 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat
     7981 cccctatggt cgactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg
     8041 ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca
     8101 aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gg
//
