LOCUS       V019765                 9118 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019765
VERSION     V019765
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             2297..3226
                     /label="SIRT5(NM_012241)"
                     /note="SIRT5(NM_012241)"
                     /gene="SIRT5"
     promoter        3277..3488
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3501..3769
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      3798..3807
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             3828..4490
                     /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             4560..4613
                     /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             4614..5213
                     /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    5214..5801
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5875..6108
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6180..6301
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6320..6455
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6493..6509)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6517..6533
                     /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(6541..6571)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6586..6607
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6895..7483)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7654..8514)
                     /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(8515..8619)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9093..9109
                     /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 tagcgaattc gccaccatgc gacctctcca gattgtccca agtcgattga tttcccagct
     2341 atattgtggc ctgaagcctc cagcgtccac acgaaaccag atttgcctga aaatggctcg
     2401 gccaagttca agtatggcag attttcgaaa gttttttgca aaagcaaagc acatagtcat
     2461 catctcagga gctggtgtta gtgcagaaag tggtgttccg accttcagag gagctggagg
     2521 ttattggaga aaatggcaag cccaggacct ggcgactccc ctggcctttg cccacaaccc
     2581 gtcccgggtg tgggagttct accactaccg gcgggaggtc atggggagca aggagcccaa
     2641 cgccgggcac cgcgccatag ccgagtgtga gacccggctg ggcaagcagg gccggcgagt
     2701 cgtggtcatc acccagaaca tcgatgagct gcaccgcaag gctggcacca agaaccttct
     2761 ggagatccat ggtagcttat ttaaaactcg atgtacctct tgtggagttg tggctgagaa
     2821 ttacaagagt ccaatttgtc cagctttatc aggaaaaggt gctccagaac ctggaactca
     2881 agatgccagc atcccagttg agaaacttcc ccggtgtgaa gaggcaggct gcgggggctt
     2941 gctgcgacct cacgtcgtgt ggtttggaga aaacctggat cctgccattc tggaggaggt
     3001 tgacagagag ctcgcccact gtgatttatg tctagtggtg ggcacttcct ctgtggtgta
     3061 cccagcagcc atgtttgccc cccaggtggc tgccaggggc gtgccagtgg ctgaatttaa
     3121 cacggagacc accccagcta cgaacagatt caggtttcat ttccagggac cctgtggaac
     3181 gactcttcct gaagcccttg cctgtcatga aaatgaaact gtttcttaag gatccgcggc
     3241 cgcgaaggat ctgcgatcgc tccggtgccc gtcagtgggc agagcgcaca tcgcccacag
     3301 tccccgagaa gttgggggga ggggtcggca attgaacggg tgcctagaga aggtggcgcg
     3361 gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag
     3421 aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg tttgccgcca
     3481 gaacacagct gaagcttcga ggggctcgca tctctccttc acgcgcccgc cgccctacct
     3541 gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc tcccgcctgt ggtgcctcct
     3601 gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc gagaccgggc ctttgtccgg
     3661 cgctcccttg gagcctacct agactcagcc ggctctccac gctttgcctg accctgcttg
     3721 ctcaactcta cgtctttgtt tcgttttctg ttctgcgccg ttacagatcc aagctgtgac
     3781 cggcgcctac gctagacgcc accatggaga gcgacgagag cggcctgccc gccatggaga
     3841 tcgagtgccg catcaccggc accctgaacg gcgtggagtt cgagctggtg ggcggcggag
     3901 agggcacccc caagcagggc cgcatgacca acaagatgaa gagcaccaaa ggcgccctga
     3961 ccttcagccc ctacctgctg agccacgtga tgggctacgg cttctaccac ttcggcacct
     4021 accccagcgg ctacgagaac cccttcctgc acgccatcaa caacggcggc tacaccaaca
     4081 cccgcatcga gaagtacgag gacggcggcg tgctgcacgt gagcttcagc taccgctacg
     4141 aggccggccg cgtgatcggc gacttcaagg tggtgggcac cggcttcccc gaggacagcg
     4201 tgatcttcac cgacaagatc atccgcagca acgccaccgt ggagcacctg caccccatgg
     4261 gcgataacgt gctggtgggc agcttcgccc gcaccttcag cctgcgcgac ggcggctact
     4321 acagcttcgt ggtggacagc cacatgcact tcaagagcgc catccacccc agcatcctgc
     4381 agaacggggg ccccatgttc gccttccgcc gcgtggagga gctgcacagc aacaccgagc
     4441 tgggcatcgt ggagtaccag cacgccttca agacccccat cgccttcgcc agatcccgcg
     4501 ctcagtcgtc caattctgcc gtggacggca ccgccggacc cggctccacc ggatctcgcg
     4561 agggcagagg aagtcttcta acatgcggtg acgtggagga gaatcccggc cctatgaccg
     4621 agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc
     4681 tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca
     4741 tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca
     4801 aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg
     4861 aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc
     4921 tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt
     4981 ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg
     5041 tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct
     5101 ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg
     5161 aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgaaatcaac
     5221 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     5281 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     5341 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     5401 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     5461 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     5521 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     5581 ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg
     5641 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     5701 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt ctccgccttc
     5761 gccctcagac gagtcggatc tccctttggc cgcctccccg cctggtacct ttaagaccaa
     5821 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag
     5881 ggctaattca ctcccaacga aaataagatc tgctttttgc ttgtactggg tctctctggt
     5941 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
     6001 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
     6061 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat
     6121 gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga
     6181 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     6241 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     6301 atcatgtctg gctctagcta tcccgcccct aactccgccc agttccgccc attctccgcc
     6361 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     6421 attccagaag tagtgaggag gcttttttgg aggcctagac ttttgcagag acggcccaaa
     6481 ttcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     6541 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     6601 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     6661 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     6721 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     6781 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     6841 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     6901 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     6961 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     7021 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     7081 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     7141 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     7201 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     7261 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     7321 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     7381 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     7441 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     7501 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     7561 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     7621 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     7681 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     7741 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     7801 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     7861 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     7921 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     7981 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     8041 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     8101 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     8161 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     8221 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     8281 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     8341 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     8401 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     8461 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     8521 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     8581 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     8641 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     8701 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     8761 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     8821 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     8881 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     8941 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     9001 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
     9061 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg ccaagctg
//
