LOCUS       V020532                 8926 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020532
VERSION     V020532
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    1798..1915
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2005..2208
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2289..3035
                     /label="TSPAN7(NM_004615)"
                     /note="TSPAN7(NM_004615)"
                     /gene="TSPAN7"
     promoter        3085..3296
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3309..3577
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      3606..3615
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             3636..4298
                     /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             4368..4421
                     /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             4422..5021
                     /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    5022..5609
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5683..5916
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5988..6109
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6128..6263
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6301..6317)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6325..6341
                     /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(6349..6379)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6394..6415
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6703..7291)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7462..8322)
                     /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(8323..8427)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8901..8917
                     /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 ggttaacttt
     1801 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc
     1861 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttatcga
     1921 tactagtatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     1981 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
     2041 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     2101 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
     2161 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac
     2221 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag attctagagc
     2281 tagccaccat ggcatcgagg agaatggaga ccaaacctgt gataacctgt ctcaaaaccc
     2341 tcctcatcat ctactccttc gtcttctgga tcactggggt gatcctgctg gctgttggag
     2401 tctggggcaa acttactctg ggcacctata tctcccttat tgccgagaac tccacaaatg
     2461 ctccctatgt gctcatcgga actggcacca ctattgttgt ctttggcctg tttggatgct
     2521 ttgctacatg tcgtggtagc ccatggatgc tgaaactgta tgccatgttt ctgtccctgg
     2581 tgttcctggc tgagctcgta gctggcattt cagggtttgt gtttcgtcat gagatcaagg
     2641 acaccttcct gaggacttac acggacgcta tgcagactta caatggcaat gatgagagga
     2701 gccgggcagt ggaccatgtg cagcgcagcc tgagctgctg tggtgtgcag aactacacca
     2761 actggagcac cagcccctac ttcctggagc atggcatccc ccccagctgc tgcatgaacg
     2821 aaactgattg taatccccag gatctacaca atctgactgt ggccgccacc aaagttaacc
     2881 agaagggttg ttatgatctg gtaactagtt tcatggagac taacatggga atcatcgctg
     2941 gagtggcgtt tggaatcgca ttctcccagt taattggcat gctgctggcc tgctgtctgt
     3001 cccggttcat cacggccaat cagtatgaga tggtgtaagg atccgcggcc gcaaggatct
     3061 gcgatcgctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt
     3121 tggggggagg ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg gtaaactggg
     3181 aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa
     3241 gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacagctga
     3301 agcttcgagg ggctcgcatc tctccttcac gcgcccgccg ccctacctga ggccgccatc
     3361 cacgccggtt gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga actgcgtccg
     3421 ccgtctaggt aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg ctcccttgga
     3481 gcctacctag actcagccgg ctctccacgc tttgcctgac cctgcttgct caactctacg
     3541 tctttgtttc gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg gcgcctacgc
     3601 tagacgccac catggagagc gacgagagcg gcctgcccgc catggagatc gagtgccgca
     3661 tcaccggcac cctgaacggc gtggagttcg agctggtggg cggcggagag ggcaccccca
     3721 agcagggccg catgaccaac aagatgaaga gcaccaaagg cgccctgacc ttcagcccct
     3781 acctgctgag ccacgtgatg ggctacggct tctaccactt cggcacctac cccagcggct
     3841 acgagaaccc cttcctgcac gccatcaaca acggcggcta caccaacacc cgcatcgaga
     3901 agtacgagga cggcggcgtg ctgcacgtga gcttcagcta ccgctacgag gccggccgcg
     3961 tgatcggcga cttcaaggtg gtgggcaccg gcttccccga ggacagcgtg atcttcaccg
     4021 acaagatcat ccgcagcaac gccaccgtgg agcacctgca ccccatgggc gataacgtgc
     4081 tggtgggcag cttcgcccgc accttcagcc tgcgcgacgg cggctactac agcttcgtgg
     4141 tggacagcca catgcacttc aagagcgcca tccaccccag catcctgcag aacgggggcc
     4201 ccatgttcgc cttccgccgc gtggaggagc tgcacagcaa caccgagctg ggcatcgtgg
     4261 agtaccagca cgccttcaag acccccatcg ccttcgccag atcccgcgct cagtcgtcca
     4321 attctgccgt ggacggcacc gccggacccg gctccaccgg atctcgcgag ggcagaggaa
     4381 gtcttctaac atgcggtgac gtggaggaga atcccggccc tatgaccgag tacaagccca
     4441 cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt
     4501 tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca
     4561 ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg
     4621 cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg
     4681 tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc
     4741 aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca
     4801 ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg
     4861 gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca
     4921 acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag
     4981 gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg aaatcaacct ctggattaca
     5041 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     5101 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     5161 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     5221 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     5281 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     5341 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     5401 tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga
     5461 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     5521 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct ccgccttcgc cctcagacga
     5581 gtcggatctc cctttggccg cctccccgcc tggtaccttt aagaccaatg acttacaagg
     5641 cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact
     5701 cccaacgaaa ataagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct
     5761 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     5821 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     5881 tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta
     5941 ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg
     6001 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     6061 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc
     6121 tctagctatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     6181 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     6241 gtgaggaggc ttttttggag gcctagactt ttgcagagac ggcccaaatt cgtaatcatg
     6301 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     6361 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     6421 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     6481 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     6541 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     6601 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     6661 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     6721 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     6781 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     6841 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     6901 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     6961 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     7021 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     7081 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     7141 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     7201 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     7261 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     7321 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     7381 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     7441 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     7501 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     7561 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     7621 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     7681 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     7741 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     7801 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     7861 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     7921 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     7981 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     8041 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     8101 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     8161 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     8221 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     8281 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     8341 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     8401 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     8461 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     8521 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     8581 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     8641 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     8701 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     8761 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     8821 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     8881 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctg
//
