LOCUS       V027101                 9630 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027101
VERSION     V027101
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     LTR             234..414
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /label="5' LTR (truncated)"
     misc_feature    458..583
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1076..1309
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1493..1537
                     /codon_start=1
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /product="antigenic peptide corresponding to amino acids
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
                     et al., 2013)"
                     /transl_table=1
                     /translation="KNEQELLELDKWASL"
                     /label="gp41 peptide"
     misc_feature    1798..1915
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2005..2208
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2297..3817
                     /label="Cgas(NM_173386)"
                     /note="Cgas(NM_173386)"
                     /gene="Cgas"
     CDS             3839..4558
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK*"
                     /label="EGFP"
     promoter        4600..4811
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4824..5092
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5119..5718
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     misc_feature    5725..6313
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6387..6620
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6692..6813
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6832..6967
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7005..7021)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7029..7045
                     /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(7053..7083)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7098..7119
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7407..7995)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8166..9026)
                     /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(9027..9131)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9605..9621
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
ORIGIN
        1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
       61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
      121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
      181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc
      241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
      421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct
      481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
      541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
      601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
      661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
      721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
      781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
      841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
      901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg
      961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc
     1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     1381 aatctctgga acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggttaacttt
     1801 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc
     1861 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttatcga
     1921 tactagtatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     1981 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
     2041 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     2101 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
     2161 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac
     2221 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag attctagagc
     2281 tagcgaattc gccaccatgg aagatccgcg tagaaggacg acggcgccac gcgctaagaa
     2341 gccgtccgcg aagcgcgccc cgacgcagcc cagcaggacc agggcccacg cggaaagctg
     2401 cggcccgcaa aggggggctc gatcgcggcg ggcggagcgt gacggggaca ccacggagaa
     2461 gccacgtgcc ccagggcccc gagtgcatcc agcaagggcc actgagctca ccaaagatgc
     2521 acagccctcg gccatggacg cggcaggagc caccgcgcgg cctgccgtcc gggtgcccca
     2581 gcagcaggcc atcctggatc cggagctgcc cgccgtacgg gagccccagc cgcccgcgga
     2641 tcccgaggcg cggaaagtcg taaggggacc tagccacaga aggggcgcgc gctccaccgg
     2701 gcagcccaga gcgccgcgag ggtccaggaa ggaaccggac aagctaaaga aggtgctgga
     2761 caaattgaga ttgaaacgca aagatatctc ggaggcggcc gagacggtga ataaagttgt
     2821 ggaacgcctg ctgcgcagaa tgcagaaacg ggagtcggag ttcaaaggtg tggagcagct
     2881 gaacactggc agctactatg aacatgtgaa gatttctgct cctaatgaat ttgatgttat
     2941 gtttaaactg gaagtcccca ggattgagct acaagaatat tatgaaacag gtgctttcta
     3001 tcttgtgaaa ttcaaaagaa ttccacgagg aaatccgctg agtcatttct tagaagggga
     3061 agtattatca gctaccaaga tgctgtcaaa gtttaggaaa atcattaaag aagaagttaa
     3121 agaaatcaaa gatatagatg tcagtgtgga gaaggaaaaa ccaggaagcc ctgctgtaac
     3181 acttcttatc aggaaccctg aagaaatctc tgtggatata attctggctt tggagtcaaa
     3241 aggcagctgg cctattagta ccaaagaagg actacctatt caaggctggc tgggcacaaa
     3301 agtgaggacc aatctaagac gagagccgtt ttatctcgta cccaagaatg caaaggatgg
     3361 aaatagtttt caaggagaga cctggcgcct ctctttctct cacactgaaa agtacatttt
     3421 gaataatcac gggatagaga aaacatgctg tgaatcttcc ggagcaaaat gctgcagaaa
     3481 agaatgttta aaattaatga aatacctttt ggaacagttg aaaaaagagt ttcaagagct
     3541 ggatgcattc tgttcctacc atgtgaaaac tgccatcttt cacatgtgga cccaggaccc
     3601 gcaggacagt cagtgggacc ccaggaacct cagctcctgc ttcgataagt tgttagcatt
     3661 ctttcttgag tgcctcagga cagagaaact ggatcattat tttattccaa agttcaatct
     3721 attctctcaa gaactaattg accgaaaaag taaagaattt ctatcgaaga aaattgaata
     3781 tgaaagaaat aatgggtttc caatttttga caagcttgga tccggaggag ggggaagcat
     3841 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     3901 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     3961 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     4021 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
     4081 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     4141 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     4201 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     4261 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     4321 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     4381 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     4441 cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     4501 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaagc
     4561 ggccgcgaag gatctgcgat cgctccggtg cccgtcagtg ggcagagcgc acatcgccca
     4621 cagtccccga gaagttgggg ggaggggtcg gcaattgaac gggtgcctag agaaggtggc
     4681 gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg
     4741 gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg
     4801 ccagaacaca gctgaagctt cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta
     4861 cctgaggccg ccatccacgc cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct
     4921 cctgaactgc gtccgccgtc taggtaagtt taaagctcag gtcgagaccg ggcctttgtc
     4981 cggcgctccc ttggagccta cctagactca gccggctctc cacgctttgc ctgaccctgc
     5041 ttgctcaact ctacgtcttt gtttcgtttt ctgttctgcg ccgttacaga tccaagctgt
     5101 gaccggcgcc tacgctagat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac
     5161 gacgtccccc gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc
     5221 cacaccgtcg acccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc
     5281 acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg
     5341 gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc
     5401 atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg
     5461 ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac
     5521 cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc
     5581 ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc
     5641 ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc
     5701 cgcaagcccg gtgcctgagt cgacaatcaa cctctggatt acaaaatttg tgaaagattg
     5761 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     5821 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     5881 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     5941 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     6001 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     6061 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
     6121 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     6181 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     6241 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     6301 gccgcctccc cgcctggtac ctttaagacc aatgacttac aaggcagctg tagatcttag
     6361 ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac gaaaataaga
     6421 tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc
     6481 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
     6541 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
     6601 agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt atttataact
     6661 tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt ataatggtta
     6721 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     6781 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc tatcccgccc
     6841 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     6901 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     6961 ggaggcctag acttttgcag agacggccca aattcgtaat catggtcata gctgtttcct
     7021 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     7081 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     7141 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     7201 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     7261 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     7321 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     7381 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     7441 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     7501 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     7561 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     7621 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     7681 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     7741 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     7801 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     7861 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     7921 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     7981 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     8041 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     8101 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     8161 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     8221 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     8281 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     8341 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     8401 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     8461 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     8521 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     8581 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     8641 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     8701 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     8761 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     8821 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     8881 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     8941 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     9001 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     9061 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     9121 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     9181 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt
     9241 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     9301 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     9361 ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt
     9421 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc gccattcagg
     9481 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg
     9541 aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
     9601 cgttgtaaaa cgacggccag tgccaagctg
//
