LOCUS       V022838                 6611 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022838
VERSION     V022838
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             941..2101
                     /label="SIRPG(NM_018556)"
                     /note="SIRPG(NM_018556)"
                     /gene="SIRPG"
     CDS             2108..2173
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2211..2435
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2481..2909
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2923..3252
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3319..4113
                     /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    4287..4408
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4457..4473)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4481..4497
                     /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(4505..4535)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4550..4571
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4859..5444)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5615..6475)
                     /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(6476..6580)
                     /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 atccgccacc atgcctgtcc cagcctcctg
      961 gccccatcct cctggtcctt tcctgcttct gactctactg ctgggactta cagaagtggc
     1021 aggtgaggag gagctacaga tgattcagcc tgagaagctc ctgttggtca cagttggaaa
     1081 gacagccact ctgcactgca ctgtgacctc cctgcttccc gtgggacccg tcctgtggtt
     1141 cagaggagtt ggaccaggcc gggaattaat ctacaatcaa aaagaaggcc acttccccag
     1201 ggtaacaaca gtttcagacc tcacaaagag aaacaacatg gacttttcca tccgcatcag
     1261 tagcatcacc ccagcagatg tcggcacata ctactgtgtg aagtttcgaa aagggagccc
     1321 tgagaacgtg gagtttaagt ctggaccagg cactgagatg gctttgggtg ccaaaccctc
     1381 tgcccccgtg gtattgggcc ctgcggcgag gaccacacct gagcatacag tgagtttcac
     1441 ctgtgagtcc catggcttct ctcccagaga catcaccctg aaatggttca aaaatgggaa
     1501 tgagctctca gacttccaga ccaacgtgga ccccacagga cagagtgtgg cctacagcat
     1561 ccgcagcaca gccagggtgg tactggaccc ctgggacgtt cgctctcagg tcatctgcga
     1621 ggtggcccat gtcaccttgc agggggaccc tcttcgtggg actgccaact tgtctgaggc
     1681 catccgagtt ccacccacct tggaggttac tcaacagccc atgagggtgg ggaaccaggt
     1741 aaacgtcacc tgccaggtga ggaagttcta cccccagagc ctacagctga cctggtcgga
     1801 gaatggaaac gtgtgccaga gagaaacagc ctcgaccctt acagagaaca aggatggtac
     1861 ctacaactgg acaagctggt tcctggtgaa catatctgac caaagggatg atgtggtcct
     1921 cacctgccag gtgaagcatg atgggcagct ggcggtcagc aaacgccttg ccctagaggt
     1981 cacagtccac cagaaggacc agagctcaga tgctacccct ggcccggcat catcccttac
     2041 tgcgctgctc ctcatagctg tcctcctggg ccccatctac gtcccctgga agcagaagac
     2101 cctcgaggac tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga
     2161 tgacgatgac aagtagtgag ggcccgttta aacccgctga tcagcctcga ctgtgccttc
     2221 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     2281 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     2341 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     2401 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     2461 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     2521 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     2581 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     2641 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     2701 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     2761 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     2821 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     2881 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     2941 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3001 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3061 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     3121 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     3181 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     3241 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca
     3301 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     3361 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     3421 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     3481 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     3541 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     3601 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     3661 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     3721 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     3781 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     3841 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     3901 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     3961 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     4021 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     4081 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     4141 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     4201 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     4261 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     4321 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     4381 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     4441 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     4501 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     4561 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     4621 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     4681 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     4741 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     4801 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     4861 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     4921 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     4981 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     5041 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     5101 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     5161 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     5221 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     5281 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     5341 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggttttt
     5401 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     5461 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     5521 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     5581 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     5641 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     5701 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     5761 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     5821 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     5881 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     5941 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     6001 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     6061 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     6121 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     6181 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     6241 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     6301 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     6361 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     6421 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     6481 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     6541 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     6601 cacctgacgt c
//
