LOCUS       V040961                 9493 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V040961
VERSION     V040961
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..3519
                     /label="CRTAP(NM_006371)"
                     /note="CRTAP(NM_006371)"
                     /gene="CRTAP"
     CDS             3526..3591
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3635..3846
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3859..4127
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      4156..4165
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             4186..4848
                     /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             4918..4971
                     /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             4972..5571
                     /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    5572..6160
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6234..6467
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6539..6660
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6700..6835
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6868..6884)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6892..6908
                     /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(6916..6946)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6961..6982
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7270..7858)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8029..8889)
                     /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(8890..8994)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9468..9484
                     /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 gccaccatgg agccggggcg ccggggggcc
     2341 gcggcgctgc tagcgctgct gtgcgtggcc tgcgcgctgc gcgccgggcg cgcccaatac
     2401 gaacgctaca gcttccgcag cttcccacgg gacgagctga tgccgctcga gtcggcctac
     2461 cggcacgcgc tggacaagta cagcggcgag cactgggccg agagcgtggg ctacctggag
     2521 atcagcctgc ggctgcaccg cttgctgcgc gacagcgagg ccttctgcca ccgcaactgc
     2581 agcgccgcgc cgcagcccga gcccgccgcc ggcctcgcca gctatcccga gctgcgcctc
     2641 ttcgggggcc tgctgcgccg cgcgcactgc ctcaagcgct gcaagcaggg cctgccagcc
     2701 ttccgccagt cccagcccag ccgcgaggtg ctggcggact tccagcgccg cgagccctac
     2761 aagttcctgc agttcgctta cttcaaggca aataatctcc ccaaagccat cgccgctgct
     2821 cacacctttc tactgaagca tcctgatgac gaaatgatga agaggaacat ggcatattat
     2881 aagagcctgc ctggtgccga ggactacatt aaagacctgg aaaccaagtc atatgaaagc
     2941 ctgttcatcc gagcagtgcg ggcatacaac ggtgagaact ggagaacatc catcacagac
     3001 atggagctgg cccttcccga cttcttcaaa gccttttacg agtgtctcgc agcctgcgag
     3061 ggttccaggg agatcaagga cttcaaggat ttctaccttt ccatagcaga tcattatgta
     3121 gaagttctgg aatgcaaaat acagtgtgaa gagaacctca ccccagttat aggaggctat
     3181 ccggttgaga aatttgtggc taccatgtat cattacttgc agtttgccta ttataagttg
     3241 aacgacctga agaatgcagc cccctgtgca gtcagctatc tgctctttga tcagaatgac
     3301 aaggtcatgc agcagaacct ggtgtattac cagtaccaca gggacacttg gggcctctcg
     3361 gatgagcact tccagcccag acctgaagca gttcagttct ttaatgtgac cacactccag
     3421 aaggagctgt atgactttgc taaggaaaat ataatggatg atgatgaggg agaagttgtg
     3481 gaatatgtgg atgacctctt ggaactggag gagaccagcg gatccgacta caaagaccat
     3541 gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa gtagcggccg
     3601 cgaaggatct gcgatcgctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc
     3661 cccgagaagt tggggggagg ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg
     3721 gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa
     3781 ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga
     3841 acacagctga agcttcgagg ggctcgcatc tctccttcac gcgcccgccg ccctacctga
     3901 ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga
     3961 actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg
     4021 ctcccttgga gcctacctag actcagccgg ctctccacgc tttgcctgac cctgcttgct
     4081 caactctacg tctttgtttc gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg
     4141 gcgcctacgc tagacgccac catggagagc gacgagagcg gcctgcccgc catggagatc
     4201 gagtgccgca tcaccggcac cctgaacggc gtggagttcg agctggtggg cggcggagag
     4261 ggcaccccca agcagggccg catgaccaac aagatgaaga gcaccaaagg cgccctgacc
     4321 ttcagcccct acctgctgag ccacgtgatg ggctacggct tctaccactt cggcacctac
     4381 cccagcggct acgagaaccc cttcctgcac gccatcaaca acggcggcta caccaacacc
     4441 cgcatcgaga agtacgagga cggcggcgtg ctgcacgtga gcttcagcta ccgctacgag
     4501 gccggccgcg tgatcggcga cttcaaggtg gtgggcaccg gcttccccga ggacagcgtg
     4561 atcttcaccg acaagatcat ccgcagcaac gccaccgtgg agcacctgca ccccatgggc
     4621 gataacgtgc tggtgggcag cttcgcccgc accttcagcc tgcgcgacgg cggctactac
     4681 agcttcgtgg tggacagcca catgcacttc aagagcgcca tccaccccag catcctgcag
     4741 aacgggggcc ccatgttcgc cttccgccgc gtggaggagc tgcacagcaa caccgagctg
     4801 ggcatcgtgg agtaccagca cgccttcaag acccccatcg ccttcgccag atcccgcgct
     4861 cagtcgtcca attctgccgt ggacggcacc gccggacccg gctccaccgg atctcgcgag
     4921 ggcagaggaa gtcttctaac atgcggtgac gtggaggaga atcccggccc tatgaccgag
     4981 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc
     5041 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc
     5101 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag
     5161 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa
     5221 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg
     5281 gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg
     5341 ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc
     5401 gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc
     5461 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag
     5521 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg aaatcaacct
     5581 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     5641 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     5701 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     5761 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     5821 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     5881 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     5941 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt
     6001 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     6061 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     6121 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggtacctt taagaccaat
     6181 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg
     6241 gctaattcac tcccaacgaa gataagatct gctttttgct tgtactgggt ctctctggtt
     6301 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
     6361 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
     6421 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg
     6481 tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa
     6541 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     6601 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     6661 tcatgtctgg ctctagctat cccgccccta actccgccca tcccgcccct aactccgccc
     6721 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     6781 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctagac
     6841 ttttgcagag accaaattcg taatcatgtc atagctgttt cctgtgtgaa attgttatcc
     6901 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     6961 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     7021 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     7081 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     7141 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     7201 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     7261 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     7321 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     7381 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     7441 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     7501 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     7561 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     7621 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     7681 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     7741 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     7801 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     7861 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     7921 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     7981 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     8041 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     8101 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     8161 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     8221 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     8281 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     8341 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     8401 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     8461 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     8521 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     8581 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     8641 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     8701 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     8761 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     8821 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     8881 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     8941 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     9001 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     9061 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     9121 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     9181 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     9241 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
     9301 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga
     9361 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
     9421 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
     9481 cagtgccaag ctg
//
