LOCUS       V019321                 8636 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019321
VERSION     V019321
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             2291..3388
                     /label="Setd7(NM_080793)"
                     /note="Setd7(NM_080793)"
                     /gene="Setd7"
     promoter        3439..3650
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3663..3931
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      3960..3969
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             3990..4652
                     /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"
     misc_feature    4731..5319
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5393..5626
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5698..5819
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      5838..5973
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6011..6027)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6035..6051
                     /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(6059..6089)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6104..6125
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6413..7001)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7172..8032)
                     /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(8033..8137)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8611..8627
                     /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 tagcgccacc atggatagcg acgatgaggt ggtggaggaa gccgtggaag ggcacctgga
     2341 cgatgacgga ttaccacacg ggttctgcac tgtcacctac tcctccacgg acagatttga
     2401 gggcaacttt gtgcatggag aaaagaatgg gcgtgggaaa ttcttcttct ttgacggcag
     2461 caccctggag ggatattacg tggacgatgc cctgcagggc cagggcgttt acacctatga
     2521 ggatggaggt gttctccaag gcacctatgt ggatggagag ctgaatggac cggcccagga
     2581 gtacgactct gacgggaggc tgatcttcaa gggacagtac aaagacaaca atcgacacgg
     2641 agtgtgttgg atccattacc cggatggagg cagcctcgtc ggggaagtca atgaagatgg
     2701 agagatgacg ggagaaaaga tcgcctatgt gtaccctgac cagaggactg cgctctatgg
     2761 gaagttcatt gacggagaga tgctcgaagg gaaactggcc accctcatgg ccaccgaaga
     2821 agggaggcca cactttgaag tgacgtcggg aagttccgtg taccactttg ataaatcgac
     2881 ttcctcctgc atctccagcg atgctctcct tccagaccct tatgagtctg agagggttta
     2941 tgttgccgac tctctcatct ccagtgctgg agaagggctg ttttccaagg tagcagttgg
     3001 acctaatact gttatgtcgt tttataatgg agtccgaatt acacaccaag aggttgacag
     3061 cagggattgg gccctaaatg ggaacactct ctccctggac gaggagacag tcattgatgt
     3121 gcccgagccc tacaaccacg tgtccaagta ctgtgcctcc ttgggacaca aggccaatca
     3181 ctccttcact ccgaactgcg tctatgacct gtttgtccac ccccgtttcg ggccgatcaa
     3241 gtgcatccgc accctgcgag ctgtggaggc cgaggaagag ctgaccgttg cctacggcta
     3301 tgaccacagc cccccgggga agagtgggcc tgaagccccc gagtggtatc aagtggagct
     3361 gaaggccttc caggccaccc agcagaagtg aggatccgcg gccgcgaagg atctgcgatc
     3421 gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg
     3481 gaggggtcgg caattgaacg ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg
     3541 atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag
     3601 tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag ctgaagcttc
     3661 gaggggctcg catctctcct tcacgcgccc gccgccctac ctgaggccgc catccacgcc
     3721 ggttgagtcg cgttctgccg cctcccgcct gtggtgcctc ctgaactgcg tccgccgtct
     3781 aggtaagttt aaagctcagg tcgagaccgg gcctttgtcc ggcgctccct tggagcctac
     3841 ctagactcag ccggctctcc acgctttgcc tgaccctgct tgctcaactc tacgtctttg
     3901 tttcgttttc tgttctgcgc cgttacagat ccaagctgtg accggcgcct acgctagacg
     3961 ccaccatgga gagcgacgag agcggcctgc ccgccatgga gatcgagtgc cgcatcaccg
     4021 gcaccctgaa cggcgtggag ttcgagctgg tgggcggcgg agagggcacc cccaagcagg
     4081 gccgcatgac caacaagatg aagagcacca aaggcgccct gaccttcagc ccctacctgc
     4141 tgagccacgt gatgggctac ggcttctacc acttcggcac ctaccccagc ggctacgaga
     4201 accccttcct gcacgccatc aacaacggcg gctacaccaa cacccgcatc gagaagtacg
     4261 aggacggcgg cgtgctgcac gtgagcttca gctaccgcta cgaggccggc cgcgtgatcg
     4321 gcgacttcaa ggtggtgggc accggcttcc ccgaggacag cgtgatcttc accgacaaga
     4381 tcatccgcag caacgccacc gtggagcacc tgcaccccat gggcgataac gtgctggtgg
     4441 gcagcttcgc ccgcaccttc agcctgcgcg acggcggcta ctacagcttc gtggtggaca
     4501 gccacatgca cttcaagagc gccatccacc ccagcatcct gcagaacggg ggccccatgt
     4561 tcgccttccg ccgcgtggag gagctgcaca gcaacaccga gctgggcatc gtggagtacc
     4621 agcacgcctt caagaccccc atcgccttcg ccagatcccg cgctcagtcg tccaattctg
     4681 ccgtggacgg caccgccgga cccggctcca ccggatctcg ctaagtcgac aatcaacctc
     4741 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     4801 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     4861 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
     4921 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
     4981 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
     5041 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     5101 acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg
     5161 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
     5221 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc
     5281 ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggtaccttt aagaccaatg
     5341 acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg
     5401 ctaattcact cccaacgaaa ataagatctg ctttttgctt gtactgggtc tctctggtta
     5461 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     5521 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     5581 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt
     5641 catcttatta ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggaac
     5701 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     5761 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     5821 catgtctggc tctagctatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     5881 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     5941 tccagaagta gtgaggaggc ttttttggag gcctagactt ttgcagagac ggcccaaatt
     6001 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     6061 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     6121 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     6181 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     6241 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     6301 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     6361 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     6421 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     6481 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     6541 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     6601 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     6661 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     6721 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     6781 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     6841 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     6901 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     6961 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     7021 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     7081 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     7141 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     7201 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     7261 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     7321 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     7381 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     7441 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     7501 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     7561 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     7621 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     7681 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     7741 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     7801 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     7861 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     7921 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     7981 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     8041 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     8101 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     8161 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     8221 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     8281 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     8341 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
     8401 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     8461 gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     8521 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     8581 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctg
//
