LOCUS       V044773                 7752 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044773
VERSION     V044773
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..2432
                     /label="St7l(NM_153091)"
                     /note="St7l(NM_153091)"
                     /gene="St7l"
     CDS             2439..2504
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2528..3116
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3182..3230
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3432..3860
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3874..4203
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4270..5064
                     /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    5240..5361
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5410..5426)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5434..5450
                     /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(5458..5488)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5503..5524
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5812..6400)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6571..7431)
                     /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(7432..7536)
                     /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 ccaccatggc ggacggagac tacttcgcgg
      781 aagccgcgcg ggccgtgggg tgtccccatg cacctgttcc tggcttgggc ctgggcccgc
      841 cgctgggctg gaaagagcgg ctgaaggccg ggttggcgaa ctctgggtcc acgctgtggt
      901 tcctggcggg gctggggctg ctttacgctc tgagggtccc gctgaggctg tgtgacaacg
      961 tgacagcggt gacagggttt ttaagttcac tgacacccaa gttctatgtg gcacttacag
     1021 ggacatcttc tttgatatct ggactaatat ttatatttga atggtggtac ttccataagc
     1081 atggtacatc ttttattgaa caagtgtcca taagccattt gcggccactg atgggaggaa
     1141 cagagagtag catttcagag ccgggctctc ctgccaacag cagagaaagt gaaaccctca
     1201 gacaccacca tctatcagaa tgcaaggtct ggagaaaccc tttaaacctc ttcagaggag
     1261 cagaatacag aagatacact tgggtgactg gtaaagagcc acttacttac tatgacatga
     1321 acttgtctgc ccaggatcac cagacctttt tcacctgtga aacagacttt ttacgtcctt
     1381 cagacacagt tatgcagaag gcatggaggg aaagaaatcc tccagctcga atcaaagcag
     1441 cctatcaagc tttagagtta aacaatgact gtgccactgc ctacgtccta ctggccgaag
     1501 aagaagcaac aaccattgta gatgctgaaa ggttgtttaa acaggcactc agggcaggag
     1561 agataattta tagacggtca cagcaatgcc agcaccagag ccctcagcat gaagctcaac
     1621 tgaggcgaga taccaatgta cttgtgtata ttaaaagaag attggccatg tgtgcgagaa
     1681 aattgggaag aataagagaa gcagtgaaaa taatgagaga tttgatgaaa gaatttcctc
     1741 ctcttaccat gttgaacatc catgaaaatc ttttagaatc acttctagaa ttgcaggcct
     1801 atgcagatgt tcaggcagtc ctggcaaaat atgatgatat aagccttcca aagtcagcgg
     1861 ccatctgcta cacagcagca ctactgaaaa caagaaccgt ctcagataaa ttctctccag
     1921 aaacagcctt cagaaaaggg ttaagcacag cagaaattaa tgctgtggaa gcaattcata
     1981 gagctgtgga atttaatcct catgttccaa aatacttact agagatgaaa agtttaatct
     2041 tacctccaga acacattctg aagcgaggag atagcgaagc aattgcctat gctttctttc
     2101 atctacagca ctggaaacga atagaaggtg ctctgaatct cctgcagtgt acatgggaag
     2161 gcacttttag aatgattcca tacccattag agaaaggaca tctattttac ccttatccca
     2221 gctgcacaga gactgctgat agagagctat tgcctacttt ccatcacgtc tctgtctatc
     2281 caaagaagga gcttcctttt ttcatccact ttacagcagg actgtgctcc tctaccgcaa
     2341 tgctagcgtt tctcactcac cagtttcctg aagtcatggg tgtttttgct aaagctgtga
     2401 gtatgatctc caggacttgt gtggagtatt taggatccga ctacaaagac catgacggtg
     2461 attataaaga tcatgacatc gactacaagg atgacgatga caagtagtga gggcccgata
     2521 tctcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     2581 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     2641 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     2701 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     2761 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     2821 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     2881 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg
     2941 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     3001 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     3061 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg
     3121 aaacggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     3181 acggcaataa aaagacagaa taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg
     3241 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     3301 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     3361 cgcagccaac gtcggggcgg caggccctgc catagcagat ctgcgcagct ggggctctag
     3421 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     3481 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     3541 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcggggca tccctttagg
     3601 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     3661 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     3721 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     3781 ttttgattta taagggattt tggggatttc ggcctattgg ttaaaaaatg agctgattta
     3841 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     3901 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     3961 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4021 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     4081 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     4141 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     4201 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     4261 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     4321 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     4381 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     4441 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     4501 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     4561 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     4621 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     4681 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     4741 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     4801 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     4861 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     4921 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     4981 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     5041 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgcgaaatga ccgaccaagc
     5101 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     5161 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     5221 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     5281 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     5341 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     5401 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     5461 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     5521 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     5581 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     5641 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     5701 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     5761 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     5821 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     5881 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     5941 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     6001 ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     6061 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     6121 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     6181 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     6241 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     6301 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     6361 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     6421 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     6481 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     6541 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     6601 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     6661 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     6721 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     6781 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     6841 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     6901 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     6961 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     7021 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     7081 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     7141 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     7201 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     7261 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     7321 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     7381 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     7441 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     7501 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     7561 tgacgtcgac ggatcgggag atctcccgat cccctatggt cgactctcag tacaatctgc
     7621 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
     7681 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
     7741 aatctgctta gg
//
