LOCUS       V024493                 9154 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024493
VERSION     V024493
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..3268
                     /label="SFXN1(NM_022754)"
                     /note="SFXN1(NM_022754)"
                     /gene="SFXN1"
     promoter        3313..3524
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3537..3805
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      3834..3843
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             3864..4526
                     /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             4596..4649
                     /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             4650..5249
                     /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    5250..5837
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5911..6144
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6216..6337
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6356..6491
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6529..6545)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6553..6569
                     /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(6577..6607)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6622..6643
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6931..7519)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7690..8550)
                     /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(8551..8655)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9129..9145
                     /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 ccatgtctgg agaactacca ccaaacatta acatcaagga
     2341 acctcgatgg gatcaaagca ctttcattgg acgagccaat catttcttca ctgtaactga
     2401 ccccaggaac attctgttaa ccaacgaaca actcgagagt gcgagaaaaa tagtacatga
     2461 ttacaggcaa ggaattgttc ctcctggtct tacagaaaat gaattgtgga gagcaaagta
     2521 catctatgat tcagcttttc atcctgacac tggtgagaag atgattttga taggaagaat
     2581 gtcagcccag gttcccatga acatgaccat cacaggttgt atgatgacgt tttacaggac
     2641 tacgccggct gtgctgttct ggcagtggat taaccagtcc ttcaatgccg tcgtcaatta
     2701 caccaacaga agtggagacg cacccctcac tgtcaatgag ttgggaacag cttacgtttc
     2761 tgcaacaact ggtgccgtag caacagctct aggactcaat gcattgacca agcatgtctc
     2821 accactgata ggacgttttg ttccctttgc tgccgtagct gctgctaatt gcattaatat
     2881 tccattaatg aggcaaaggg aactcaaagt tggcattccc gtcacggatg agaatgggaa
     2941 ccgcttgggg gagtcggcga acgctgcgaa acaagccatc acgcaagttg tcgtgtccag
     3001 gattctcatg gcagcccctg gcatggccat ccctccattc attatgaaca ctttggaaaa
     3061 gaaagccttt ttgaagaggt tcccatggat gagtgcaccc attcaagttg ggttagttgg
     3121 cttctgtttg gtgtttgcta cacccctgtg ttgtgccctg tttcctcaga aaagttccat
     3181 gtctgtgaca agcttggagg ccgagttgca agctaagatc caagagagcc atcctgaatt
     3241 gcgacgcgtg tacttcaata agggattgta agcggccgcg aaggatctgc gatcgctccg
     3301 gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg gggggagggg
     3361 tcggcaattg aacgggtgcc tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg
     3421 tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt gcagtagtcg
     3481 ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acagctgaag cttcgagggg
     3541 ctcgcatctc tccttcacgc gcccgccgcc ctacctgagg ccgccatcca cgccggttga
     3601 gtcgcgttct gccgcctccc gcctgtggtg cctcctgaac tgcgtccgcc gtctaggtaa
     3661 gtttaaagct caggtcgaga ccgggccttt gtccggcgct cccttggagc ctacctagac
     3721 tcagccggct ctccacgctt tgcctgaccc tgcttgctca actctacgtc tttgtttcgt
     3781 tttctgttct gcgccgttac agatccaagc tgtgaccggc gcctacgcta gacgccacca
     3841 tggagagcga cgagagcggc ctgcccgcca tggagatcga gtgccgcatc accggcaccc
     3901 tgaacggcgt ggagttcgag ctggtgggcg gcggagaggg cacccccaag cagggccgca
     3961 tgaccaacaa gatgaagagc accaaaggcg ccctgacctt cagcccctac ctgctgagcc
     4021 acgtgatggg ctacggcttc taccacttcg gcacctaccc cagcggctac gagaacccct
     4081 tcctgcacgc catcaacaac ggcggctaca ccaacacccg catcgagaag tacgaggacg
     4141 gcggcgtgct gcacgtgagc ttcagctacc gctacgaggc cggccgcgtg atcggcgact
     4201 tcaaggtggt gggcaccggc ttccccgagg acagcgtgat cttcaccgac aagatcatcc
     4261 gcagcaacgc caccgtggag cacctgcacc ccatgggcga taacgtgctg gtgggcagct
     4321 tcgcccgcac cttcagcctg cgcgacggcg gctactacag cttcgtggtg gacagccaca
     4381 tgcacttcaa gagcgccatc caccccagca tcctgcagaa cgggggcccc atgttcgcct
     4441 tccgccgcgt ggaggagctg cacagcaaca ccgagctggg catcgtggag taccagcacg
     4501 ccttcaagac ccccatcgcc ttcgccagat cccgcgctca gtcgtccaat tctgccgtgg
     4561 acggcaccgc cggacccggc tccaccggat ctcgcgaggg cagaggaagt cttctaacat
     4621 gcggtgacgt ggaggagaat cccggcccta tgaccgagta caagcccacg gtgcgcctcg
     4681 ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc
     4741 ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag
     4801 aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg
     4861 ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga
     4921 tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag
     4981 gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct
     5041 cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg
     5101 ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct
     5161 acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct
     5221 ggtgcatgac ccgcaagccc ggtgcctgaa atcaacctct ggattacaaa atttgtgaaa
     5281 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     5341 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     5401 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     5461 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     5521 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     5581 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     5641 ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     5701 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     5761 tgccggctct gcggcctctt ccgcgtctcc gccttcgccc tcagacgagt cggatctccc
     5821 tttggccgcc tccccgcctg gtacctttaa gaccaatgac ttacaaggca gctgtagatc
     5881 ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaaaat
     5941 aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc
     6001 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
     6061 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
     6121 agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat
     6181 aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg
     6241 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     6301 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc
     6361 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     6421 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     6481 ttttggaggc ctagactttt gcagagacgg cccaaattcg taatcatggt catagctgtt
     6541 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     6601 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     6661 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     6721 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     6781 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     6841 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     6901 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     6961 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     7021 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     7081 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     7141 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     7201 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     7261 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     7321 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     7381 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     7441 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     7501 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     7561 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     7621 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     7681 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     7741 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     7801 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     7861 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     7921 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     7981 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     8041 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     8101 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     8161 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     8221 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     8281 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     8341 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     8401 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     8461 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     8521 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     8581 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     8641 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     8701 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     8761 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     8821 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     8881 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     8941 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt
     9001 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     9061 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     9121 acgacgttgt aaaacgacgg ccagtgccaa gctg
//
