LOCUS       V030386                 9086 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030386
VERSION     V030386
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..3180
                     /label="PDCD1(NM_005018)"
                     /note="PDCD1(NM_005018)"
                     /gene="PDCD1"
     promoter        3245..3456
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3469..3737
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      3766..3775
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             3796..4458
                     /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             4528..4581
                     /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             4582..5181
                     /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    5182..5769
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5843..6076
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6148..6269
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6288..6423
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6461..6477)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6485..6501
                     /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(6509..6539)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6554..6575
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6863..7451)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7622..8482)
                     /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(8483..8587)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9061..9077
                     /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 gccaccatgc agatcccaca ggcgccctgg
     2341 ccagtcgtct gggcggtgct acaactgggc tggcggccag gatggttctt agactcccca
     2401 gacaggccct ggaacccccc caccttctcc ccagccctgc tcgtggtgac cgaaggggac
     2461 aacgccacct tcacctgcag cttctccaac acatcggaga gcttcgtgct aaactggtac
     2521 cgcatgagcc ccagcaacca gacggacaag ctggccgcct tccccgagga ccgcagccag
     2581 cccggccagg actgccgctt ccgtgtcaca caactgccca acgggcgtga cttccacatg
     2641 agcgtggtca gggcccggcg caatgacagc ggcacctacc tctgtggggc catctccctg
     2701 gcccccaagg cgcagatcaa agagagcctg cgggcagagc tcagggtgac agagagaagg
     2761 gcagaagtgc ccacagccca ccccagcccc tcacccaggc cagccggcca gttccaaacc
     2821 ctggtggttg gtgtcgtggg cggcctgctg ggcagcctgg tgctgctagt ctgggtcctg
     2881 gccgtcatct gctcccgggc cgcacgaggg acaataggag ccaggcgcac cggccagccc
     2941 ctgaaggagg acccctcagc cgtgcctgtg ttctctgtgg actatgggga gctggatttc
     3001 cagtggcgag agaagacccc ggagcccccc gtgccctgtg tccctgagca gacggagtat
     3061 gccaccattg tctttcctag cggaatgggc acctcatccc ccgcccgcag gggctcagct
     3121 gacggccctc ggagtgccca gccactgagg cctgaggatg gacactgctc ttggcccctc
     3181 tgaggatccc tgcgatatcc agcgcggccg cgaaggatct gcgatcgctc cggtgcccgt
     3241 cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat
     3301 tgaacgggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg
     3361 ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac
     3421 gttctttttc gcaacgggtt tgccgccaga acacagctga agcttcgagg ggctcgcatc
     3481 tctccttcac gcgcccgccg ccctacctga ggccgccatc cacgccggtt gagtcgcgtt
     3541 ctgccgcctc ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt aagtttaaag
     3601 ctcaggtcga gaccgggcct ttgtccggcg ctcccttgga gcctacctag actcagccgg
     3661 ctctccacgc tttgcctgac cctgcttgct caactctacg tctttgtttc gttttctgtt
     3721 ctgcgccgtt acagatccaa gctgtgaccg gcgcctacgc tagacgccac catggagagc
     3781 gacgagagcg gcctgcccgc catggagatc gagtgccgca tcaccggcac cctgaacggc
     3841 gtggagttcg agctggtggg cggcggagag ggcaccccca agcagggccg catgaccaac
     3901 aagatgaaga gcaccaaagg cgccctgacc ttcagcccct acctgctgag ccacgtgatg
     3961 ggctacggct tctaccactt cggcacctac cccagcggct acgagaaccc cttcctgcac
     4021 gccatcaaca acggcggcta caccaacacc cgcatcgaga agtacgagga cggcggcgtg
     4081 ctgcacgtga gcttcagcta ccgctacgag gccggccgcg tgatcggcga cttcaaggtg
     4141 gtgggcaccg gcttccccga ggacagcgtg atcttcaccg acaagatcat ccgcagcaac
     4201 gccaccgtgg agcacctgca ccccatgggc gataacgtgc tggtgggcag cttcgcccgc
     4261 accttcagcc tgcgcgacgg cggctactac agcttcgtgg tggacagcca catgcacttc
     4321 aagagcgcca tccaccccag catcctgcag aacgggggcc ccatgttcgc cttccgccgc
     4381 gtggaggagc tgcacagcaa caccgagctg ggcatcgtgg agtaccagca cgccttcaag
     4441 acccccatcg ccttcgccag atcccgcgct cagtcgtcca attctgccgt ggacggcacc
     4501 gccggacccg gctccaccgg atctcgcgag ggcagaggaa gtcttctaac atgcggtgac
     4561 gtggaggaga atcccggccc tatgaccgag tacaagccca cggtgcgcct cgccacccgc
     4621 gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg
     4681 cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc
     4741 ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg
     4801 gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg
     4861 cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg
     4921 gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac
     4981 caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc
     5041 gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg
     5101 ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg
     5161 acccgcaagc ccggtgcctg aaatcaacct ctggattaca aaatttgtga aagattgact
     5221 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     5281 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     5341 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     5401 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     5461 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     5521 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg
     5581 acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     5641 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     5701 ctgcggcctc ttccgcgtct ccgccttcgc cctcagacga gtcggatctc cctttggccg
     5761 cctccccgcc tggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac
     5821 tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaaa ataagatctg
     5881 ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc
     5941 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
     6001 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
     6061 tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt ataacttgca
     6121 aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa
     6181 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     6241 ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa
     6301 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     6361 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     6421 gcctagactt ttgcagagac ggcccaaatt cgtaatcatg gtcatagctg tttcctgtgt
     6481 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     6541 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     6601 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     6661 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     6721 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     6781 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     6841 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     6901 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     6961 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     7021 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     7081 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     7141 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     7201 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     7261 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     7321 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     7381 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     7441 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     7501 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     7561 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     7621 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     7681 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     7741 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     7801 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     7861 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     7921 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     7981 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     8041 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     8101 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     8161 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     8221 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     8281 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     8341 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     8401 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     8461 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     8521 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     8581 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     8641 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg
     8701 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
     8761 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct
     8821 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa
     8881 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc
     8941 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag
     9001 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
     9061 gtaaaacgac ggccagtgcc aagctg
//
