LOCUS       V033654                 9224 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033654
VERSION     V033654
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             2317..3318
                     /label="Jun(NM_010591)"
                     /note="Jun(NM_010591)"
                     /gene="Jun"
     promoter        3383..3594
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3607..3875
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      3904..3913
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             3934..4596
                     /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             4666..4719
                     /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             4720..5319
                     /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    5320..5907
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5981..6214
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6286..6407
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6426..6561
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6599..6615)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6623..6639
                     /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(6647..6677)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6692..6713
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7001..7589)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7760..8620)
                     /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(8621..8725)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9199..9215
                     /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 ctgatacgaa ctcggaattc gccaccatga ctgcaaagat ggaaacgacc
     2341 ttctacgacg atgccctcaa cgcctcgttc ctccagtccg agagcggtgc ctacggctac
     2401 agtaacccta agatcctaaa acagagcatg accttgaacc tggccgaccc ggtgggcagt
     2461 ctgaagccgc acctccgcgc caagaactcg gaccttctca cgtcgcccga cgtcgggctg
     2521 ctcaagctgg cgtcgccgga gctggagcgc ctgatcatcc agtccagcaa tgggcacatc
     2581 accactacac cgacccccac ccagttcttg tgccccaaga acgtgaccga cgagcaggag
     2641 ggcttcgccg agggcttcgt gcgcgccctg gctgaactgc atagccagaa cacgcttccc
     2701 agtgtcacct ccgcggcaca gccggtcagc ggggcgggca tggtggctcc cgcggtggcc
     2761 tcagtagcag gcgctggcgg cggtggtggc tacagcgcca gcctgcacag tgagcctccg
     2821 gtctacgcca acctcagcaa cttcaacccg ggtgcgctga gcagcggcgg tggggcgccc
     2881 tcctatggcg cggccgggct ggcctttccc tcgcagccgc agcagcagca gcagccgcct
     2941 cagccgccgc accacttgcc ccaacagatc ccggtgcagc acccgcggct gcaagccctg
     3001 aaggaagagc cgcagaccgt gccggagatg ccgggagaga cgccgcccct gtcccctatc
     3061 gacatggagt ctcaggagcg gatcaaggca gagaggaagc gcatgaggaa ccgcattgcc
     3121 gcctccaagt gccggaaaag gaagctggag cggatcgctc ggctagagga aaaagtgaaa
     3181 accttgaaag cgcaaaactc cgagctggca tccacggcca acatgctcag ggaacaggtg
     3241 gcacagctta agcagaaagt catgaaccac gttaacagtg ggtgccaact catgctaacg
     3301 cagcagttgc aaacgttttg aggatccctg cgatatccag cgcggccgcg aaggatctgc
     3361 gatcgctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg
     3421 gggggagggg tcggcaattg aacgggtgcc tagagaaggt ggcgcggggt aaactgggaa
     3481 agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt
     3541 gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acagctgaag
     3601 cttcgagggg ctcgcatctc tccttcacgc gcccgccgcc ctacctgagg ccgccatcca
     3661 cgccggttga gtcgcgttct gccgcctccc gcctgtggtg cctcctgaac tgcgtccgcc
     3721 gtctaggtaa gtttaaagct caggtcgaga ccgggccttt gtccggcgct cccttggagc
     3781 ctacctagac tcagccggct ctccacgctt tgcctgaccc tgcttgctca actctacgtc
     3841 tttgtttcgt tttctgttct gcgccgttac agatccaagc tgtgaccggc gcctacgcta
     3901 gacgccacca tggagagcga cgagagcggc ctgcccgcca tggagatcga gtgccgcatc
     3961 accggcaccc tgaacggcgt ggagttcgag ctggtgggcg gcggagaggg cacccccaag
     4021 cagggccgca tgaccaacaa gatgaagagc accaaaggcg ccctgacctt cagcccctac
     4081 ctgctgagcc acgtgatggg ctacggcttc taccacttcg gcacctaccc cagcggctac
     4141 gagaacccct tcctgcacgc catcaacaac ggcggctaca ccaacacccg catcgagaag
     4201 tacgaggacg gcggcgtgct gcacgtgagc ttcagctacc gctacgaggc cggccgcgtg
     4261 atcggcgact tcaaggtggt gggcaccggc ttccccgagg acagcgtgat cttcaccgac
     4321 aagatcatcc gcagcaacgc caccgtggag cacctgcacc ccatgggcga taacgtgctg
     4381 gtgggcagct tcgcccgcac cttcagcctg cgcgacggcg gctactacag cttcgtggtg
     4441 gacagccaca tgcacttcaa gagcgccatc caccccagca tcctgcagaa cgggggcccc
     4501 atgttcgcct tccgccgcgt ggaggagctg cacagcaaca ccgagctggg catcgtggag
     4561 taccagcacg ccttcaagac ccccatcgcc ttcgccagat cccgcgctca gtcgtccaat
     4621 tctgccgtgg acggcaccgc cggacccggc tccaccggat ctcgcgaggg cagaggaagt
     4681 cttctaacat gcggtgacgt ggaggagaat cccggcccta tgaccgagta caagcccacg
     4741 gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc
     4801 gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc
     4861 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg
     4921 gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg
     4981 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa
     5041 cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc
     5101 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga
     5161 gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac
     5221 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga
     5281 ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgaa atcaacctct ggattacaaa
     5341 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     5401 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     5461 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     5521 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     5581 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     5641 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     5701 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt
     5761 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     5821 cgcggcctgc tgccggctct gcggcctctt ccgcgtctcc gccttcgccc tcagacgagt
     5881 cggatctccc tttggccgcc tccccgcctg gtacctttaa gaccaatgac ttacaaggca
     5941 gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc
     6001 caacgaaaat aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga
     6061 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
     6121 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
     6181 agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt
     6241 cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca
     6301 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     6361 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc
     6421 tagctatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     6481 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     6541 gaggaggctt ttttggaggc ctagactttt gcagagacgg cccaaattcg taatcatggt
     6601 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     6661 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     6721 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     6781 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     6841 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     6901 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     6961 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     7021 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     7081 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     7141 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     7201 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     7261 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     7321 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     7381 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     7441 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     7501 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     7561 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     7621 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     7681 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     7741 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     7801 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     7861 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     7921 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     7981 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     8041 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     8101 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     8161 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     8221 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     8281 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     8341 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     8401 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     8461 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     8521 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     8581 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     8641 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     8701 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     8761 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     8821 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     8881 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     8941 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     9001 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
     9061 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     9121 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
     9181 tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gctg
//
