LOCUS       V043251                10102 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043251
VERSION     V043251
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..1920
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2010..2213
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2302..4956
                     /label="KMT5B(NM_017635)"
                     /note="KMT5B(NM_017635)"
                     /gene="KMT5B"
     CDS             4963..5028
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        5072..5283
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             5296..5564
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5591..6190
                     /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    6197..6785
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6859..7092
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    7164..7285
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      7304..7439
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7477..7493)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7501..7517
                     /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(7525..7555)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7570..7591
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7879..8467)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8638..9498)
                     /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(9499..9603)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     10077..10093
                     /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 attacaaaat tcaaaatttt
     1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag
     2221 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct
     2281 agagctagcg aattcgccac catgaagtgg ttgggagaat ccaagaacat ggtggtgaat
     2341 ggcaggagaa atggaggcaa gttgtctaat gaccatcagc agaatcaatc aaaattacag
     2401 cacacgggga aggacaccct gaaggctggc aaaaatgcag tcgagaggag gtcgaacaga
     2461 tgtaatggta actcgggatt tgaaggacag agtcgctatg taccatcctc tggaatgtcc
     2521 gccaaggaac tctgtgaaaa tgatgaccta gcaaccagtt tggttcttga tccctattta
     2581 ggttttcaaa cacacaaaat gaatactagc gcctttcctt cgaggagctc aaggcatttt
     2641 tcaaaatctg acagtttttc tcacaacaac cctgtgagat ttaggcctat taaaggaagg
     2701 caggaagaac taaaggaagt aattgaacgt tttaagaaag atgaacactt ggagaaagcc
     2761 ttcaaatgtt tgacttcagg cgaatgggca cggcactatt ttctcaacaa gaataaaatg
     2821 caggagaaat tattcaaaga acatgtattt atttatttgc gaatgtttgc aactgacagt
     2881 ggatttgaaa tattgccatg taatagatac tcatcagaac aaaatggagc caaaatagtt
     2941 gcaacaaaag agtggaaacg aaatgacaaa atagaattac tggtgggttg tattgccgaa
     3001 ctttcagaaa ttgaggagaa catgctactt agacatggag aaaacgactt cagtgtcatg
     3061 tactccacaa ggaaaaactg tgctcaactc tggctgggtc ctgctgcgtt tataaaccat
     3121 gattgcagac ctaattgtaa gtttgtgtca actggtcgag atacagcatg tgtgaaggct
     3181 ctaagagaca ttgaacctgg agaagaaatt tcttgttatt atggagatgg gttctttgga
     3241 gaaaataatg agttctgcga gtgttacact tgcgaaagac ggggcactgg tgcttttaaa
     3301 tccagagtgg gactgcctgc gcctgctcct gttatcaata gcaaatatgg actcagagaa
     3361 acagataaac gtttaaatag gcttaaaaag ttaggtgaca gcagcaaaaa ttcagacagt
     3421 caatctgtca gctctaacac tgatgcagat accactcagg aaaaaaacaa tgcaacttct
     3481 aaccgaaaat cttcagttgg cgtaaaaaag aatagcaaga gcagaacgtt aacgaggcaa
     3541 tctatgtcaa gaattccagc ttcttccaac tctacctcat ctaagctaac tcatataaat
     3601 aattccaggg taccaaagaa actgaagaag cctgcaaagc ctttactttc aaagataaaa
     3661 ttgagaaatc attgcaagcg gctggagcaa aagaatgctt caagaaaact cgaaatggga
     3721 aacttagtac tgaaagagcc taaagtagtt ctgtataaaa atttgcccat taaaaaagat
     3781 aaggagccag agggaccagc ccaagccgca gttgccagcg ggtgcttgac tagacacgcg
     3841 gcgagagaac acagacagaa tcctgtgaga ggtgctcatt cgcaggggga gagctcgccc
     3901 tgcacctaca taactcggcg gtcagtgagg acaagaacaa atctgaagga ggcctctgac
     3961 atcaagcttg aaccaaatac gttgaatggc tataaaagca gtgtgacgga accttgcccc
     4021 gacagtggtg aacagctgca gccagctcct gtgctgcagg aggaagaact ggctcatgag
     4081 actgcacaaa aaggggaggc aaagtgtcat aagagtgaca caggcatgtc caaaaagaag
     4141 tcacgacaag gaaaacttgt gaaacagttt gcaaaaatag aggaatctac tccagtgcac
     4201 gattctcctg gaaaagacga cgcggtacca gatttgatgg gtccccattc tgaccagggt
     4261 gagcacagtg gcactgtggg cgtgcctgtg agctacacag actgtgctcc ttcacccgtc
     4321 ggttgttcag ttgtgacatc agatagcttc aaaacaaaag acagctttag aactgcaaaa
     4381 agtaaaaaga agaggcgaat cacaaggtat gatgcacagt taatcctaga aaataactct
     4441 gggattccca aattgactct tcgtaggcgt catgatagca gcagcaaaac aaatgaccaa
     4501 gagaatgatg gaatgaactc ttccaaaata agcatcaagt taagcaaaga ccatgacaac
     4561 gataacaatc tctatgtagc aaagcttaat aatggattta actcaggatc aggcagtagt
     4621 tctacaaaat taaaaatcca gctaaaacga gatgaggaaa atagggggtc ttatacagag
     4681 gggcttcatg aaaatggggt gtgctgcagt gatcctcttt ctctcttgga gtctcgaatg
     4741 gaggtggatg actatagtca gtatgaggaa gaaagtacag atgattcctc ctcttctgag
     4801 ggcgatgaag aggaggatga ctatgatgat gactttgaag acgattttat tcctcttcct
     4861 ccagctaagc gcttgaggtt aatagttgga aaagactcta tagatattga catttcttca
     4921 aggagaagag aagatcagtc tttaaggctt aatgccggat ccgactacaa agaccatgac
     4981 ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta gcggccgcga
     5041 aggatctgcg atcgctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc
     5101 gagaagttgg ggggaggggt cggcaattga acgggtgcct agagaaggtg gcgcggggta
     5161 aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg
     5221 tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca
     5281 cagctgaagc ttcgaggggc tcgcatctct ccttcacgcg cccgccgccc tacctgaggc
     5341 cgccatccac gccggttgag tcgcgttctg ccgcctcccg cctgtggtgc ctcctgaact
     5401 gcgtccgccg tctaggtaag tttaaagctc aggtcgagac cgggcctttg tccggcgctc
     5461 ccttggagcc tacctagact cagccggctc tccacgcttt gcctgaccct gcttgctcaa
     5521 ctctacgtct ttgtttcgtt ttctgttctg cgccgttaca gatccaagct gtgaccggcg
     5581 cctacgctag atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc
     5641 ccgggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt
     5701 cgacccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt
     5761 cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac
     5821 cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga
     5881 gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg
     5941 gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa
     6001 gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc
     6061 cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac
     6121 cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc
     6181 cggtgcctga gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     6241 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     6301 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     6361 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     6421 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     6481 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     6541 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
     6601 ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     6661 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     6721 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     6781 cccgcctggt acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt
     6841 taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaaaataa gatctgcttt
     6901 ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
     6961 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
     7021 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
     7081 aaatctctag cagtagtagt tcatgtcatc ttattattca gtatttataa cttgcaaaga
     7141 aatgaatatc agagagtgag aggaacttgt ttattgcagc ttataatggt tacaaataaa
     7201 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     7261 tgtccaaact catcaatgta tcttatcatg tctggctcta gctatcccgc ccctaactcc
     7321 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     7381 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     7441 agacttttgc agagacggcc caaattcgta atcatggtca tagctgtttc ctgtgtgaaa
     7501 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     7561 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     7621 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     7681 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     7741 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     7801 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     7861 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     7921 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     7981 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     8041 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     8101 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     8161 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     8221 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     8281 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     8341 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     8401 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     8461 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     8521 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     8581 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     8641 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     8701 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     8761 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     8821 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     8881 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     8941 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     9001 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     9061 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     9121 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     9181 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     9241 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     9301 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     9361 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     9421 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     9481 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     9541 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     9601 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     9661 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     9721 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     9781 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     9841 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
     9901 gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa
     9961 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
    10021 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
    10081 aacgacggcc agtgccaagc tg
//
