LOCUS       V044847                 8412 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044847
VERSION     V044847
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..3092
                     /label="Supv3l1(NM_181423)"
                     /note="Supv3l1(NM_181423)"
                     /gene="Supv3l1"
     CDS             3099..3164
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3188..3776
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3842..3890
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4092..4520
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4534..4863
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4930..5724
                     /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    5900..6021
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6070..6086)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6094..6110
                     /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(6118..6148)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6163..6184
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6472..7060)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7231..8091)
                     /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(8092..8196)
                     /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 ccaccatgtc cttgcctcgg tgtaccttgc
      781 tgtgggctcg gctcccggcg gggcgcgggg ctgggccccg ggcggcccct tgctccgccc
      841 ttcgcgctct cgtggggtcg ttcccggggg cttcaggacg cgttccctgc ttggccgcct
      901 cctcctctgc ctctggtggc tccaaagccc cgaacacgtc cttgttcgtg ccgctgacgg
      961 tgaagcctca gggtcccagc gccgatggcg acgtgggcgc ggagctcact cggcctctag
     1021 acaagaatga agtaaagaag atcttagaca agttttataa gaggcaagaa attcaaaagc
     1081 tgagtgctga ctatggactt gatgctcgtc tcttccacca ggctttcata agcttcagga
     1141 attacattat gcaatcccat tctctggacg tggacattca cattgtcctg aatgatatct
     1201 gcttcagtgc agctcatgtg gacgatttat ttccattttt cttgagacat gccaaacaaa
     1261 tatttcctgt attggaatgc aaggatgatc tacgtaaaat cagtgacttg aggataccac
     1321 ccaactggta cccagaagcc agagcgagac agcggaagat aatattccac tcaggcccta
     1381 ccaacagtgg gaagacttac catgccatcc agagatacct gtcagcgacc tctggcgtgt
     1441 actgcggtcc tctaaaactg ctggcgcacg agatctttga gaagagcaat gctgctggtg
     1501 tgccgtgtga cttggtgaca ggtgaagagc ggctgacagt ggagcctgag gggaaacaag
     1561 ccacccatgt ctcttgcact gtggagatgt gcaatgttgc aactccttat gaagtggcag
     1621 tgatagatga aatccagatg attcgagacc cagccagagg atgggcttgg accagagcac
     1681 tgctagggct ctgtgccgag gaagtccatt tatgtggaga gtctgctgct attaacctgg
     1741 tctctgagct cctgtacacc actggggagg aagtagaggt tcagaagtat gagaggctta
     1801 cccccatttc tgtgctggat catgcgctgg agtctttgga caaccttcag cctggggact
     1861 gcattgtctg ctttagcaag aatgacattt attctgtgag ccgacagatt gaaattcggg
     1921 gactggaatc cgcggtgata tacggcagct tgccacctgg gaccaaactt gctcaagcaa
     1981 gaaagtttaa tgaccccaat gatccatgca agatcctggt agccacagat gcaattggca
     2041 tgggacttaa tttgagcata aggagaatta ttttttactc ccttataaag cccagcatca
     2101 atgaaaaggg agagaaggag ctggagccca tcaccacctc ccaagccctg cagattgcag
     2161 gcagagctgg cagattcagc tcacacttta aagaggggca ggttaccaca atgcaccgag
     2221 acgacctggc tttattgaag gacattttga atagacctgt ggatcctata caggccgccg
     2281 gtctccaccc gactgccgag cagattgaga tgtttgccta ccacctcccc gagacaacgc
     2341 tgtccaatct cattgatatt tttgtagact ttgcacaagt tgatgggcag tactttgtct
     2401 gcaatatgga tgattttaag ttctctgcag agctgatcca gcatatccca ctaagtctcc
     2461 gggtgaggta tgtgttctgc acagccccca tcaataagaa gcagcctttt gtctgctcgt
     2521 cgttgttaca gtttgccagg cagtacagca ggaatgagcc cctgaccttt gcatggctac
     2581 gccggtacat caagtggcct ctgcttccgc ctaagaatat taaagacctc atggatcttg
     2641 aagctgtcca tgacgtcttt gatctttacc tgtggctaag ctaccgattt attgatatgt
     2701 ttccagactc tagccttgtt cgaagtctcc agaaagagct ggatgccatt atccaagaag
     2761 gcgtgcacaa catcaccaag ctgattaaaa tctctgagtc acgcaagctt ttgaatctgg
     2821 agagcttacc atcgggggac cagtcacggt tgtcaggagc ctccaagagc ccagcgagaa
     2881 ggacacgtgg caccaaaagt gcagggaata aggccacaga gccactcagc cccagtgaca
     2941 aggagctgcc ccttgcctct aggttggtgc agcaaggact cctcactgca gacatgctca
     3001 ggcagctcca gaaggagtgg ctgacacagc agccagaaca cagcagagag aaagtgggga
     3061 cacggagaaa gaagaaggac cccgactccg atggatccga ctacaaagac catgacggtg
     3121 attataaaga tcatgacatc gactacaagg atgacgatga caagtagtga gggcccgata
     3181 tctcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     3241 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     3301 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     3361 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     3421 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     3481 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     3541 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg
     3601 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     3661 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     3721 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg
     3781 aaacggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     3841 acggcaataa aaagacagaa taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg
     3901 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     3961 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     4021 cgcagccaac gtcggggcgg caggccctgc catagcagat ctgcgcagct ggggctctag
     4081 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     4141 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     4201 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcggggca tccctttagg
     4261 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     4321 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     4381 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     4441 ttttgattta taagggattt tggggatttc ggcctattgg ttaaaaaatg agctgattta
     4501 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     4561 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     4621 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4681 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     4741 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     4801 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     4861 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     4921 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     4981 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     5041 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     5101 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     5161 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     5221 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     5281 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     5341 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     5401 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     5461 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     5521 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     5581 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     5641 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     5701 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgcgaaatga ccgaccaagc
     5761 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     5821 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     5881 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     5941 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     6001 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     6061 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     6121 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     6181 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     6241 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     6301 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     6361 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     6421 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     6481 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     6541 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     6601 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     6661 ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     6721 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     6781 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     6841 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     6901 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     6961 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     7021 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     7081 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     7141 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     7201 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     7261 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     7321 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     7381 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     7441 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     7501 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     7561 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     7621 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     7681 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     7741 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     7801 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     7861 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     7921 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     7981 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     8041 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     8101 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     8161 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     8221 tgacgtcgac ggatcgggag atctcccgat cccctatggt cgactctcag tacaatctgc
     8281 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
     8341 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
     8401 aatctgctta gg
//
