LOCUS       V029339                 9340 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029339
VERSION     V029339
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             2397..3338
                     /label="PRSS21(NM_006799)"
                     /note="PRSS21(NM_006799)"
                     /gene="PRSS21"
     promoter        3499..3710
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3723..3991
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      4020..4029
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             4050..4712
                     /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             4782..4835
                     /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             4836..5435
                     /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    5436..6023
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6097..6330
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6402..6523
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6542..6677
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6715..6731)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6739..6755
                     /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(6763..6793)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6808..6829
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7117..7705)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7876..8736)
                     /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(8737..8841)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9315..9331
                     /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 tagaagatta
     2281 atactttcag gctgggagta gagggcagag ctcccacccc gccccgcccc cagggggcgc
     2341 cccgggcccg gcgcgagagg aggcagaggg ggcgtcaggc cgcgggagag gaggccatgg
     2401 gcgcgcgcgg ggcgctgctg ctggcgctgc tgctggctcg ggctggactc aggaagccgg
     2461 agtcgcagga ggcggcgccg ttatcaggac catgcggccg acgggtcatc acgtcgcgca
     2521 tcgtgggtgg agaggacgcc gaactcgggc gttggccgtg gcaggggagc ctgcgcctgt
     2581 gggattccca cgtatgcgga gtgagcctgc tcagccaccg ctgggcactc acggcggcgc
     2641 actgctttga aacctatagt gaccttagtg atccctccgg gtggatggtc cagtttggcc
     2701 agctgacttc catgccatcc ttctggagcc tgcaggccta ctacacccgt tacttcgtat
     2761 cgaatatcta tctgagccct cgctacctgg ggaattcacc ctatgacatt gccttggtga
     2821 agctgtctgc acctgtcacc tacactaaac acatccagcc catctgtctc caggcctcca
     2881 catttgagtt tgagaaccgg acagactgct gggtgactgg ctgggggtac atcaaagagg
     2941 atgaggcact gccatctccc cacaccctcc aggaagttca ggtcgccatc ataaacaact
     3001 ctatgtgcaa ccacctcttc ctcaagtaca gtttccgcaa ggacatcttt ggagacatgg
     3061 tttgtgctgg caatgcccaa ggcgggaagg atgcctgctt cggtgactca ggtggaccct
     3121 tggcctgtaa caagaatgga ctgtggtatc agattggagt cgtgagctgg ggagtgggct
     3181 gtggtcggcc caatcggccc ggtgtctaca ccaatatcag ccaccacttt gagtggatcc
     3241 agaagctgat ggcccagagt ggcatgtccc agccagaccc ctcctggccg ctactctttt
     3301 tccctcttct ctgggctctc ccactcctgg ggccggtctg agcctacctg agcccatgca
     3361 gcctggggcc actgccaagt caggccctgg ttctcttctg tcttgtttgg taataaacac
     3421 attccagttg atgccttgca gggcattctt caagtaagaa caactgaagg atctgcgatc
     3481 gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg
     3541 gaggggtcgg caattgaacg ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg
     3601 atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag
     3661 tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag ctgaagcttc
     3721 gaggggctcg catctctcct tcacgcgccc gccgccctac ctgaggccgc catccacgcc
     3781 ggttgagtcg cgttctgccg cctcccgcct gtggtgcctc ctgaactgcg tccgccgtct
     3841 aggtaagttt aaagctcagg tcgagaccgg gcctttgtcc ggcgctccct tggagcctac
     3901 ctagactcag ccggctctcc acgctttgcc tgaccctgct tgctcaactc tacgtctttg
     3961 tttcgttttc tgttctgcgc cgttacagat ccaagctgtg accggcgcct acgctagacg
     4021 ccaccatgga gagcgacgag agcggcctgc ccgccatgga gatcgagtgc cgcatcaccg
     4081 gcaccctgaa cggcgtggag ttcgagctgg tgggcggcgg agagggcacc cccaagcagg
     4141 gccgcatgac caacaagatg aagagcacca aaggcgccct gaccttcagc ccctacctgc
     4201 tgagccacgt gatgggctac ggcttctacc acttcggcac ctaccccagc ggctacgaga
     4261 accccttcct gcacgccatc aacaacggcg gctacaccaa cacccgcatc gagaagtacg
     4321 aggacggcgg cgtgctgcac gtgagcttca gctaccgcta cgaggccggc cgcgtgatcg
     4381 gcgacttcaa ggtggtgggc accggcttcc ccgaggacag cgtgatcttc accgacaaga
     4441 tcatccgcag caacgccacc gtggagcacc tgcaccccat gggcgataac gtgctggtgg
     4501 gcagcttcgc ccgcaccttc agcctgcgcg acggcggcta ctacagcttc gtggtggaca
     4561 gccacatgca cttcaagagc gccatccacc ccagcatcct gcagaacggg ggccccatgt
     4621 tcgccttccg ccgcgtggag gagctgcaca gcaacaccga gctgggcatc gtggagtacc
     4681 agcacgcctt caagaccccc atcgccttcg ccagatcccg cgctcagtcg tccaattctg
     4741 ccgtggacgg caccgccgga cccggctcca ccggatctcg cgagggcaga ggaagtcttc
     4801 taacatgcgg tgacgtggag gagaatcccg gccctatgac cgagtacaag cccacggtgc
     4861 gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg
     4921 actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc
     4981 tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg
     5041 acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg
     5101 ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga
     5161 tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg
     5221 gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg
     5281 aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc
     5341 ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc
     5401 gcacctggtg catgacccgc aagcccggtg cctgaaatca acctctggat tacaaaattt
     5461 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     5521 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     5581 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     5641 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     5701 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     5761 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     5821 tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc
     5881 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     5941 gcctgctgcc ggctctgcgg cctcttccgc gtctccgcct tcgccctcag acgagtcgga
     6001 tctccctttg gccgcctccc cgcctggtac ctttaagacc aatgacttac aaggcagctg
     6061 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac
     6121 gaaaataaga tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct
     6181 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     6241 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     6301 ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt
     6361 atttataact tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt
     6421 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     6481 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc
     6541 tatcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     6601 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     6661 aggctttttt ggaggcctag acttttgcag agacggccca aattcgtaat catggtcata
     6721 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     6781 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     6841 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     6901 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     6961 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     7021 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     7081 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     7141 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     7201 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     7261 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     7321 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     7381 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     7441 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     7501 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     7561 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     7621 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     7681 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     7741 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     7801 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     7861 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     7921 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     7981 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     8041 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     8101 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     8161 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     8221 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     8281 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     8341 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     8401 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     8461 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     8521 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     8581 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     8641 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     8701 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     8761 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     8821 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     8881 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     8941 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     9001 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     9061 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     9121 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
     9181 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     9241 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     9301 ccagtcacga cgttgtaaaa cgacggccag tgccaagctg
//
