LOCUS       V032483                 6230 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032483
VERSION     V032483
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             908..1723
                     /label="Spi1(NM_011355)"
                     /note="Spi1(NM_011355)"
                     /gene="Spi1"
     CDS             1733..1750
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     polyA_signal    1797..2021
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2067..2495
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2509..2838
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             2905..3699
                     /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    3873..3994
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4043..4059)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4067..4083
                     /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(4091..4121)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4136..4157
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4445..5030)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5201..6061)
                     /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(6062..6166)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cgccaccatg ttacaggcgt gcaaaatgga agggttttcc ctcaccgccc ctccatcgga
      961 tgacttggtt acttacgatt cagagctata ccaacgtcca atgcatgact actactcctt
     1021 cgtgggcagc gatggagaaa gccatagcga tcactactgg gatttctccg cacaccatgt
     1081 ccacaacaac gagtttgaga acttccctga gaaccacttc acagagctgc agagtgtgca
     1141 gcccccgcag ctacagcagc tctatcgcca catggagctg gaacagatgc acgtcctcga
     1201 tactcccatg gtgccacccc acaccggcct cagtcaccag gtttcctaca tgccccggat
     1261 gtgcttccct tatcaaacct tgtccccagc ccaccagcag agctcagatg aggaggaggg
     1321 tgagaggcag agccctcccc tggaggtgtc tgatggagaa gctgatggct tggagcctgg
     1381 gccaggtctt ctgcacgggg agacaggcag caagaaaaag attcgcctgt accagttcct
     1441 gctggacctg ctgcgcagcg gcgacatgaa ggacagcatc tggtgggtgg acaaggacaa
     1501 aggtaccttc cagttctcgt ccaagcacaa ggaggcgctg gcgcaccgct ggggcatcca
     1561 gaagggcaac cgcaagaaga tgacctacca gaagatggcg cgcgcgctgc gcaactacgg
     1621 caagacaggc gaggtgaaga aagtcaagaa gaagctcacc taccagttca gcggcgaggt
     1681 gctgggccgt gggggcctgg ccgagcggcg cctcccgccc cacgcggccg cacatcacca
     1741 tcatcaccat tgactcgagt ctagagggcc cgtttaaacc cgctgatcag cctcgactgt
     1801 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     1861 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     1921 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     1981 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     2041 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     2101 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     2161 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     2221 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     2281 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     2341 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     2401 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     2461 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     2521 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     2581 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     2641 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     2701 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     2761 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     2821 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     2881 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     2941 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     3001 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     3061 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     3121 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     3181 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     3241 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     3301 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     3361 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     3421 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     3481 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     3541 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     3601 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     3661 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     3721 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     3781 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     3841 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     3901 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     3961 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     4021 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     4081 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     4141 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     4201 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     4261 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     4321 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     4381 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     4441 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4501 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4561 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4621 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4681 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4741 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4801 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4861 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     4921 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4981 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     5041 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     5101 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     5161 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     5221 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     5281 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     5341 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     5401 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     5461 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     5521 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     5581 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     5641 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     5701 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     5761 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     5821 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     5881 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     5941 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     6001 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     6061 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     6121 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     6181 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
//
