LOCUS       V032083                 7328 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032083
VERSION     V032083
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..2473
                     /label="CGAS(NM_138441)"
                     /note="CGAS(NM_138441)"
                     /gene="CGAS"
     CDS             2534..2827
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /transl_table=1
                     /translation="MSDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPL
                     RRLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIGG"
                     /label="SUMO"
     CDS             2831..2848
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     polyA_signal    2895..3119
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3165..3593
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3607..3936
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4003..4797
                     /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    4971..5092
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5141..5157)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5165..5181
                     /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(5189..5219)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5234..5255
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5543..6128)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6299..7159)
                     /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(7160..7264)
                     /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 cagccttggc acggaaaggc catgcagaga gcttccgagg ccggagccac
      961 tgcccccaag gcttccgcac ggaatgccag gggcgccccg atggatccca ccgagtctcc
     1021 ggctgccccc gaggccgccc tgcctaaggc gggaaagttc ggccccgcca ggaagtcggg
     1081 atcccggcag aaaaagagcg ccccggacac ccaggagagg ccgcccgtcc gcgcaactgg
     1141 ggcccgcgcc aaaaaggccc ctcagcgcgc ccaggacacg cagccgtctg acgccaccag
     1201 cgcccctggg gcagaggggc tggagcctcc tgcggctcgg gagccggctc tttccagggc
     1261 tggttcttgc cgccagaggg gcgcgcgctg ctccacgaag ccaagacctc cgcccgggcc
     1321 ctgggacgtg cccagccccg gcctgccggt ctcggccccc attctcgtac ggagggatgc
     1381 ggcgcctggg gcctcgaagc tccgggcggt tttggagaag ttgaagctca gccgcgatga
     1441 tatctccacg gcggcgggga tggtgaaagg ggttgtggac cacctgctgc tcagactgaa
     1501 gtgcgactcc gcgttcagag gcgtcgggct gctgaacacc gggagctact atgagcacgt
     1561 gaagatttct gcacctaatg aatttgatgt catgtttaaa ctggaagtcc ccagaattca
     1621 actagaagaa tattccaaca ctcgtgcata ttactttgtg aaatttaaaa gaaatccgaa
     1681 agaaaatcct ctgagtcagt ttttagaagg tgaaatatta tcagcttcta agatgctgtc
     1741 aaagtttagg aaaatcatta aggaagaaat taacgacatt aaagatacag atgtcatcat
     1801 gaagaggaaa agaggaggga gccctgctgt aacacttctt attagtgaaa aaatatctgt
     1861 ggatataacc ctggctttgg aatcaaaaag tagctggcct gctagcaccc aagaaggcct
     1921 gcgcattcaa aactggcttt cagcaaaagt taggaagcaa ctacgactaa agccatttta
     1981 ccttgtaccc aagcatgcaa aggaaggaaa tggtttccaa gaagaaacat ggcggctatc
     2041 cttctctcac atcgaaaagg aaattttgaa caatcatgga aaatctaaaa cgtgctgtga
     2101 aaacaaagaa gagaaatgtt gcaggaaaga ttgtttaaaa ctaatgaaat accttttaga
     2161 acagctgaaa gaaaggttta aagacaaaaa acatctggat aaattctctt cttatcatgt
     2221 gaaaactgcc ttctttcacg tatgtaccca gaaccctcaa gacagtcagt gggaccgcaa
     2281 agacctgggc ctctgctttg ataactgcgt gacatacttt cttcagtgcc tcaggacaga
     2341 aaaacttgag aattatttta ttcctgaatt caatctattc tctagcaact taattgacaa
     2401 aagaagtaag gaatttctga caaagcaaat tgaatatgaa agaaacaatg agtttccagt
     2461 ttttgatgaa tttaattctg cagatatcgc caccgcggcc gcaggtggag gcggatcagg
     2521 aggcggtgga tctatgtcgg actcagaagt caatcaagaa gctaagccag aggtcaagcc
     2581 agaagtcaag cctgagactc acatcaattt aaaggtgtcc gatggatctt cagagatctt
     2641 cttcaagatc aaaaagacca ctcctttaag aaggctgatg gaagcgttcg ctaaaagaca
     2701 gggtaaggaa atggactcct taagattctt gtacgacggt attagaattc aagctgatca
     2761 gacccctgaa gatttggaca tggaggataa cgatattatt gaggctcaca gagaacagat
     2821 tggtggatca catcaccatc atcaccattg actcgagtct agagggcccg tttaaacccg
     2881 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     2941 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     3001 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     3061 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     3121 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     3181 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     3241 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     3301 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     3361 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     3421 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     3481 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     3541 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     3601 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3661 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     3721 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     3781 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     3841 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     3901 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     3961 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     4021 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     4081 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     4141 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     4201 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     4261 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     4321 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     4381 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     4441 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     4501 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     4561 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     4621 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     4681 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     4741 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     4801 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     4861 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     4921 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     4981 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     5041 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     5101 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     5161 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     5221 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     5281 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     5341 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     5401 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     5461 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     5521 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     5581 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     5641 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     5701 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     5761 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     5821 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     5881 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     5941 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     6001 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     6061 aaaccaccgc tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     6121 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     6181 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     6241 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     6301 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     6361 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     6421 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     6481 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     6541 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     6601 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     6661 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     6721 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     6781 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     6841 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     6901 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     6961 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     7021 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     7081 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     7141 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     7201 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     7261 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc
     7321 cctatggt
//
