LOCUS       V023297                 8944 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023297
VERSION     V023297
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"
     regulatory      2297..2306
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             2306..2329
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             2330..3058
                     /label="CTHRC1(NM_138455)"
                     /note="CTHRC1(NM_138455)"
                     /gene="CTHRC1"
     promoter        3103..3314
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3327..3595
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      3624..3633
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             3654..4316
                     /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             4386..4439
                     /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             4440..5039
                     /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    5040..5627
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5701..5934
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6006..6127
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6146..6281
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6319..6335)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6343..6359
                     /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(6367..6397)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6412..6433
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6721..7309)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7480..8340)
                     /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(8341..8445)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8919..8935
                     /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 gaattcgcca ccatggatta caaggatgac gacgataaga tgcgacccca
     2341 gggccccgcc gcctccccgc agcggctccg cggcctcctg ctgctcctgc tgctgcagct
     2401 gcccgcgccg tcgagcgcct ctgagatccc caaggggaag caaaaggcgc agctccggca
     2461 gagggaggtg gtggacctgt ataatggaat gtgcttacaa gggccagcag gagtgcctgg
     2521 tcgagacggg agccctgggg ccaatggcat tccgggtaca cctgggatcc caggtcggga
     2581 tggattcaaa ggagaaaagg gggaatgtct gagggaaagc tttgaggagt cctggacacc
     2641 caactacaag cagtgttcat ggagttcatt gaattatggc atagatcttg ggaaaattgc
     2701 ggagtgtaca tttacaaaga tgcgttcaaa tagtgctcta agagttttgt tcagtggctc
     2761 acttcggcta aaatgcagaa atgcatgctg tcagcgttgg tatttcacat tcaatggagc
     2821 tgaatgttca ggacctcttc ccattgaagc tataatttat ttggaccaag gaagccctga
     2881 aatgaattca acaattaata ttcatcgcac ttcttctgtg gaaggacttt gtgaaggaat
     2941 tggtgctgga ttagtggatg ttgctatctg ggttggtact tgttcagatt acccaaaagg
     3001 agatgcttct actggatgga attcagtttc tcgcatcatt attgaagaac taccaaaata
     3061 agcggccgcg aaggatctgc gatcgctccg gtgcccgtca gtgggcagag cgcacatcgc
     3121 ccacagtccc cgagaagttg gggggagggg tcggcaattg aacgggtgcc tagagaaggt
     3181 ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg
     3241 ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg
     3301 ccgccagaac acagctgaag cttcgagggg ctcgcatctc tccttcacgc gcccgccgcc
     3361 ctacctgagg ccgccatcca cgccggttga gtcgcgttct gccgcctccc gcctgtggtg
     3421 cctcctgaac tgcgtccgcc gtctaggtaa gtttaaagct caggtcgaga ccgggccttt
     3481 gtccggcgct cccttggagc ctacctagac tcagccggct ctccacgctt tgcctgaccc
     3541 tgcttgctca actctacgtc tttgtttcgt tttctgttct gcgccgttac agatccaagc
     3601 tgtgaccggc gcctacgcta gacgccacca tggagagcga cgagagcggc ctgcccgcca
     3661 tggagatcga gtgccgcatc accggcaccc tgaacggcgt ggagttcgag ctggtgggcg
     3721 gcggagaggg cacccccaag cagggccgca tgaccaacaa gatgaagagc accaaaggcg
     3781 ccctgacctt cagcccctac ctgctgagcc acgtgatggg ctacggcttc taccacttcg
     3841 gcacctaccc cagcggctac gagaacccct tcctgcacgc catcaacaac ggcggctaca
     3901 ccaacacccg catcgagaag tacgaggacg gcggcgtgct gcacgtgagc ttcagctacc
     3961 gctacgaggc cggccgcgtg atcggcgact tcaaggtggt gggcaccggc ttccccgagg
     4021 acagcgtgat cttcaccgac aagatcatcc gcagcaacgc caccgtggag cacctgcacc
     4081 ccatgggcga taacgtgctg gtgggcagct tcgcccgcac cttcagcctg cgcgacggcg
     4141 gctactacag cttcgtggtg gacagccaca tgcacttcaa gagcgccatc caccccagca
     4201 tcctgcagaa cgggggcccc atgttcgcct tccgccgcgt ggaggagctg cacagcaaca
     4261 ccgagctggg catcgtggag taccagcacg ccttcaagac ccccatcgcc ttcgccagat
     4321 cccgcgctca gtcgtccaat tctgccgtgg acggcaccgc cggacccggc tccaccggat
     4381 ctcgcgaggg cagaggaagt cttctaacat gcggtgacgt ggaggagaat cccggcccta
     4441 tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac
     4501 gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc
     4561 gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca
     4621 tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga
     4681 gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt
     4741 cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc
     4801 ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca
     4861 gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg
     4921 agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg
     4981 acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgaa
     5041 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     5101 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     5161 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     5221 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     5281 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     5341 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     5401 tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg
     5461 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     5521 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtctcc
     5581 gccttcgccc tcagacgagt cggatctccc tttggccgcc tccccgcctg gtacctttaa
     5641 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac
     5701 tggaagggct aattcactcc caacgaaaat aagatctgct ttttgcttgt actgggtctc
     5761 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
     5821 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
     5881 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta
     5941 gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg
     6001 agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     6061 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     6121 tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccagtt ccgcccattc
     6181 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     6241 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagactttt gcagagacgg
     6301 cccaaattcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     6361 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     6421 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     6481 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     6541 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     6601 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     6661 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     6721 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     6781 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     6841 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     6901 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     6961 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     7021 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     7081 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     7141 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     7201 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     7261 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     7321 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     7381 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     7441 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     7501 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     7561 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     7621 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     7681 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     7741 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     7801 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     7861 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     7921 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     7981 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     8041 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     8101 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     8161 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     8221 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     8281 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     8341 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     8401 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     8461 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     8521 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     8581 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     8641 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     8701 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
     8761 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     8821 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     8881 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa
     8941 gctg
//
