LOCUS       V039187                 9174 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V039187
VERSION     V039187
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..3372
                     /label="KMT5A(NM_020382)"
                     /note="KMT5A(NM_020382)"
                     /gene="KMT5A"
     CDS             3373..3462
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     promoter        3527..3738
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3751..4019
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      4048..4057
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             4078..4740
                     /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             4810..4863
                     /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             4864..5262
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /note="confers resistance to blasticidin"
                     /product="blasticidin S deaminase"
                     /transl_table=1
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG*"
                     /label="BSD"
     misc_feature    5269..5857
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5931..6164
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6236..6357
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6376..6511
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6549..6565)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6573..6589
                     /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(6597..6627)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6642..6663
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6951..7539)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7710..8570)
                     /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(8571..8675)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9149..9165
                     /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 ctagaggcag gaagatgtcc
     2341 aagccccgcg cggtggaggc ggcggcggcg gcggcggcgg tggcagcgac ggccccgggc
     2401 ccggagatgg tggagcggag gggcccgggg aggccccgca ccgacgggga gaacgtattt
     2461 accgggcagt caaagatcta ttcctacatg agcccgaaca aatgctctgg aatgcgtttc
     2521 ccccttcagg aagagaactc agttacacat cacgaagtca aatgccaggg gaaaccatta
     2581 gccggaatct acaggaaacg agaagagaaa agaaatgctg ggaacgcagt acggagcgcc
     2641 atgaagtccg aggaacagaa gatcaaagac gccaggaaag gtcccctggt accttttcca
     2701 aaccaaaaat ctgaagcagc agaacctcca aaaactccac cctcatcttg tgattccacc
     2761 aatgcagcca tcgccaagca agccctgaaa aagcccatca agggcaaaca ggccccccga
     2821 aaaaaagctc aaggaaaaac gcaacagaat cgcaaactta cggatttcta ccctgtccga
     2881 aggagctcca ggaagagcaa agccgagctg cagtctgaag aaaggaaaag aatagatgaa
     2941 ttgattgaaa gtgggaagga agaaggaatg aagattgacc tcatcgatgg caaaggcagg
     3001 ggtgtgattg ccaccaagca gttctcccgg ggtgactttg tggtggaata ccacggggac
     3061 ctcatcgaga tcaccgacgc caagaaacgg gaggctctgt acgcacagga cccttccacg
     3121 ggctgctaca tgtactattt tcagtatctg agcaaaacct actgcgtgga tgcaactaga
     3181 gagacaaatc gcctaggaag actgatcaat cacagcaaat gtgggaactg ccaaaccaaa
     3241 ctgcacgaca tcgacggcgt acctcacctc atcctcatcg cctcccgaga catcgcggct
     3301 ggggaggagc tcctgtatga ctatggggac cgcagcaagg cttccattga agcccacccg
     3361 tggctgaagc atgaacaaaa actaatatca gaggaagatt tggaacagaa actaatcagt
     3421 gaagaagatt tagagcaaaa gttaatttcc gaggaggact tatagggatc cctgcgatat
     3481 ccagcgcggc cgcgaaggat ctgcgatcgc tccggtgccc gtcagtgggc agagcgcaca
     3541 tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaacggg tgcctagaga
     3601 aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag
     3661 ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg
     3721 tttgccgcca gaacacagct gaagcttcga ggggctcgca tctctccttc acgcgcccgc
     3781 cgccctacct gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc tcccgcctgt
     3841 ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc gagaccgggc
     3901 ctttgtccgg cgctcccttg gagcctacct agactcagcc ggctctccac gctttgcctg
     3961 accctgcttg ctcaactcta cgtctttgtt tcgttttctg ttctgcgccg ttacagatcc
     4021 aagctgtgac cggcgcctac gctagacgcc accatggaga gcgacgagag cggcctgccc
     4081 gccatggaga tcgagtgccg catcaccggc accctgaacg gcgtggagtt cgagctggtg
     4141 ggcggcggag agggcacccc caagcagggc cgcatgacca acaagatgaa gagcaccaaa
     4201 ggcgccctga ccttcagccc ctacctgctg agccacgtga tgggctacgg cttctaccac
     4261 ttcggcacct accccagcgg ctacgagaac cccttcctgc acgccatcaa caacggcggc
     4321 tacaccaaca cccgcatcga gaagtacgag gacggcggcg tgctgcacgt gagcttcagc
     4381 taccgctacg aggccggccg cgtgatcggc gacttcaagg tggtgggcac cggcttcccc
     4441 gaggacagcg tgatcttcac cgacaagatc atccgcagca acgccaccgt ggagcacctg
     4501 caccccatgg gcgataacgt gctggtgggc agcttcgccc gcaccttcag cctgcgcgac
     4561 ggcggctact acagcttcgt ggtggacagc cacatgcact tcaagagcgc catccacccc
     4621 agcatcctgc agaacggggg ccccatgttc gccttccgcc gcgtggagga gctgcacagc
     4681 aacaccgagc tgggcatcgt ggagtaccag cacgccttca agacccccat cgccttcgcc
     4741 agatcccgcg ctcagtcgtc caattctgcc gtggacggca ccgccggacc cggctccacc
     4801 ggatctcgcg agggcagagg aagtcttcta acatgcggtg acgtggagga gaatcccggc
     4861 cctatggcca agcctttgtc tcaagaagaa tccaccctca ttgaaagagc aacggctaca
     4921 atcaacagca tccccatctc tgaagactac agcgtcgcca gcgcagctct ctctagcgac
     4981 ggccgcatct tcactggtgt caatgtatat cattttactg ggggaccttg tgcagaactc
     5041 gtggtgctgg gcactgctgc tgctgcggca gctggcaacc tgacttgtat cgtcgcgatc
     5101 ggaaatgaga acaggggcat cttgagcccc tgcggacggt gccgacaggt gcttctcgat
     5161 ctgcatcctg ggatcaaagc catagtgaag gacagtgatg gacagccgac ggcagttggg
     5221 attcgtgaat tgctgccctc tggttatgtg tgggagggct aagtcgacaa tcaacctctg
     5281 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     5341 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     5401 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     5461 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     5521 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     5581 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     5641 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc
     5701 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     5761 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     5821 cagacgagtc ggatctccct ttgggccgcc tccccgcctg gtacctttaa gaccaatgac
     5881 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct
     5941 aattcactcc caacgaaaat aagatctgct ttttgcttgt actgggtctc tctggttaga
     6001 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
     6061 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
     6121 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca
     6181 tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt
     6241 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     6301 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     6361 tgtctggctc tagctatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     6421 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     6481 cagaagtagt gaggaggctt ttttggaggc ctagactttt gcagagacgg cccaaattcg
     6541 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     6601 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     6661 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     6721 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     6781 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     6841 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     6901 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     6961 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     7021 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     7081 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     7141 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     7201 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     7261 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     7321 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     7381 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     7441 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     7501 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     7561 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     7621 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     7681 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     7741 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     7801 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     7861 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     7921 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     7981 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     8041 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     8101 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     8161 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     8221 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     8281 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     8341 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     8401 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     8461 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     8521 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     8581 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     8641 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     8701 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     8761 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     8821 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     8881 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     8941 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     9001 atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     9061 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
     9121 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gctg
//
