LOCUS       V030392                 9424 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030392
VERSION     V030392
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    1804..1921
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2011..2214
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2303..3538
                     /label="Ass1(NM_007494)"
                     /note="Ass1(NM_007494)"
                     /gene="Ass1"
     promoter        3583..3794
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3807..4075
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      4104..4113
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             4134..4796
                     /codon_start=1
                     /product="green fluorescent protein 2 from Pontellina
                     plumata, also known as ppluGFP2 (Shagin et al., 2004)"
                     /transl_table=1
                     /translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT
                     FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY
                     EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG
                     YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA"
                     /label="CopGFP"
     CDS             4866..4919
                     /codon_start=1
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding separate
                     polypeptides."
                     /product="2A peptide from&nbsp;Thosea asigna virus capsid
                     protein"
                     /protein_id=""
                     /transl_table=1
                     /translation="EGRGSLLTCGDVEENPGP"
                     /label="T2A"
     CDS             4920..5519
                     /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    5520..6107
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6181..6414
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6486..6607
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6626..6761
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6799..6815)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6823..6839
                     /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(6847..6877)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6892..6913
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7201..7789)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7960..8820)
                     /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(8821..8925)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9399..9415
                     /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 ggtatcggtt
     1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt
     1921 tatcgatact agtattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     1981 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
     2041 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
     2101 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     2161 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggttt atataagcag agctcgttta
     2221 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagattc
     2281 tagagctagc gaattcgcca ccatgtccag caagggctct gtggttctgg cctacagtgg
     2341 tggcctggac acctcctgca tcctcgtgtg gctgaaggaa caaggctatg atgtcatcgc
     2401 ctacctggcc aacattggcc agaaggaaga ctttgaggaa gccaggaaga aggcgctgaa
     2461 gcttggggcc aaaaaggtgt tcattgagga tgtgagcaag gaatttgtgg aagagttcat
     2521 ctggcctgct gtccagtcca gtgcactcta cgaggaccgc tatctcctgg gcacctctct
     2581 cgccaggcct tgcatagctc gcagacaggt ggagattgcc cagcgtgaag gggccaagta
     2641 tgtgtctcac ggcgccacgg gaaaggggaa tgaccaggtc cgctttgagc tcacctgcta
     2701 ttcactggca ccccagatta aggtcatcgc tccctggagg atgcctgagt tttacaaccg
     2761 gttcaagggc cgaaatgatc tgatggagta tgcaaagcaa cacggaatcc ccatccctgt
     2821 cacccccaag agcccctgga gtatggatga aaacctcatg cacatcagct atgaggctgg
     2881 gatcctggaa aaccccaaga atcaagcacc tccgggtctc tacacaaaaa ctcaggaccc
     2941 tgccaaagca cccaacagcc cagatgtcct tgagatagaa ttcaaaaaag gggtccctgt
     3001 gaaggtgacc aacatcaaag atggcacaac ccgcaccaca tccctggaac tcttcatgta
     3061 cctgaacgaa gttgcgggca agcacggagt gggtcgcatt gacatcgtgg agaaccgctt
     3121 cattggaatg aagtcccgag gtatctacga gaccccagca gggaccatcc tttaccacgc
     3181 tcatttagac atagaggcct tcacgatgga tcgggaagta cgcaaaatca agcagggcct
     3241 gggcctcaaa ttcgcagagc tcgtatacac aggtttctgg cacagccctg aatgtgaatt
     3301 tgttcgccac tgtatccaga agtcccagga gcgggtagaa gggaaggtgc aggtgtctgt
     3361 cttcaagggc caagtgtaca tcctcggtcg ggagtctcca ctttcactct acaatgaaga
     3421 gctggtgagc atgaacgtgc agggcgacta tgagcccatc gacgccactg gcttcatcaa
     3481 tatcaactcg ctcaggctga aggagtacca tcgccttcag agcaaggtca ctgccaaata
     3541 ggcggccgcg aaggatctgc gatcgctccg gtgcccgtca gtgggcagag cgcacatcgc
     3601 ccacagtccc cgagaagttg gggggagggg tcggcaattg aacgggtgcc tagagaaggt
     3661 ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg
     3721 ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg
     3781 ccgccagaac acagctgaag cttcgagggg ctcgcatctc tccttcacgc gcccgccgcc
     3841 ctacctgagg ccgccatcca cgccggttga gtcgcgttct gccgcctccc gcctgtggtg
     3901 cctcctgaac tgcgtccgcc gtctaggtaa gtttaaagct caggtcgaga ccgggccttt
     3961 gtccggcgct cccttggagc ctacctagac tcagccggct ctccacgctt tgcctgaccc
     4021 tgcttgctca actctacgtc tttgtttcgt tttctgttct gcgccgttac agatccaagc
     4081 tgtgaccggc gcctacgcta gacgccacca tggagagcga cgagagcggc ctgcccgcca
     4141 tggagatcga gtgccgcatc accggcaccc tgaacggcgt ggagttcgag ctggtgggcg
     4201 gcggagaggg cacccccaag cagggccgca tgaccaacaa gatgaagagc accaaaggcg
     4261 ccctgacctt cagcccctac ctgctgagcc acgtgatggg ctacggcttc taccacttcg
     4321 gcacctaccc cagcggctac gagaacccct tcctgcacgc catcaacaac ggcggctaca
     4381 ccaacacccg catcgagaag tacgaggacg gcggcgtgct gcacgtgagc ttcagctacc
     4441 gctacgaggc cggccgcgtg atcggcgact tcaaggtggt gggcaccggc ttccccgagg
     4501 acagcgtgat cttcaccgac aagatcatcc gcagcaacgc caccgtggag cacctgcacc
     4561 ccatgggcga taacgtgctg gtgggcagct tcgcccgcac cttcagcctg cgcgacggcg
     4621 gctactacag cttcgtggtg gacagccaca tgcacttcaa gagcgccatc caccccagca
     4681 tcctgcagaa cgggggcccc atgttcgcct tccgccgcgt ggaggagctg cacagcaaca
     4741 ccgagctggg catcgtggag taccagcacg ccttcaagac ccccatcgcc ttcgccagat
     4801 cccgcgctca gtcgtccaat tctgccgtgg acggcaccgc cggacccggc tccaccggat
     4861 ctcgcgaggg cagaggaagt cttctaacat gcggtgacgt ggaggagaat cccggcccta
     4921 tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac
     4981 gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc
     5041 gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca
     5101 tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga
     5161 gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt
     5221 cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc
     5281 ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca
     5341 gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg
     5401 agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg
     5461 acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgaa
     5521 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     5581 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     5641 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     5701 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     5761 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     5821 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     5881 tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg
     5941 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     6001 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtctcc
     6061 gccttcgccc tcagacgagt cggatctccc tttggccgcc tccccgcctg gtacctttaa
     6121 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac
     6181 tggaagggct aattcactcc caacgaaaat aagatctgct ttttgcttgt actgggtctc
     6241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
     6301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
     6361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta
     6421 gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg
     6481 agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     6541 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     6601 tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccagtt ccgcccattc
     6661 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     6721 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagactttt gcagagacgg
     6781 cccaaattcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     6841 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     6901 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     6961 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     7021 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     7081 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     7141 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     7201 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     7261 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     7321 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     7381 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     7441 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     7501 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     7561 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     7621 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     7681 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     7741 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     7801 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     7861 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     7921 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     7981 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     8041 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     8101 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     8161 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     8221 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     8281 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     8341 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     8401 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     8461 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     8521 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     8581 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     8641 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     8701 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     8761 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     8821 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     8881 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     8941 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     9001 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     9061 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     9121 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     9181 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
     9241 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     9301 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     9361 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa
     9421 gctg
//
