LOCUS       V023512                 9563 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023512
VERSION     V023512
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     LTR             1..634
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     misc_feature    681..806
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1303..1536
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1721..1765
                     /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    2028..2143
                     /note="central polypurine tract and central termination
                     sequence of HIV-1 (lacking the first T)"
                     /label="cPPT/CTS"
     enhancer        2201..2504
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        2505..2708
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2826..4289
                     /label="DGAT1(NM_012079)"
                     /note="DGAT1(NM_012079)"
                     /gene="DGAT1"
     CDS             4296..4319
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     promoter        4351..4850
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             4871..5470
                     /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    5484..6072
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6279..6912
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7041..7057)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7065..7081
                     /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(7089..7119)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7134..7155
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7443..8031)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8202..9062)
                     /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(9063..9167)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9215..9349
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag
     2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2161 acagcagaga tccagtttat cgataagctt gggagttccg cgttacataa cttacggtaa
     2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc
     2761 tccatagaag acaccgactc tactagagga tcgctagcgc taccggactc agatctcgag
     2821 ccaccatggg cgaccgcggc agctcccggc gccggaggac agggtcgcgg ccctcgagcc
     2881 acggcggcgg cgggcctgcg gcggcggaag aggaggtgcg ggacgccgct gcgggccccg
     2941 acgtgggagc cgcgggggac gcgccagccc cggcccccaa caaggacgga gacgccggcg
     3001 tgggcagcgg ccactgggag ctgaggtgcc atcgcctgca ggattcttta ttcagctctg
     3061 acagtggctt cagcaactac cgtggcatcc tgaactggtg tgtggtgatg ctgatcttga
     3121 gcaatgcccg gttatttctg gagaacctca tcaagtatgg catcctggtg gaccccatcc
     3181 aggtggtttc tctgttcctg aaggatccct atagctggcc cgccccatgc ctggttattg
     3241 cggccaatgt ctttgctgtg gctgcattcc aggttgagaa gcgcctggcg gtgggtgccc
     3301 tgacggagca ggcgggactg ctgctgcacg tggccaacct ggccaccatt ctgtgtttcc
     3361 cagcggctgt ggtcttactg gttgagtcta tcactccagt gggctccctg ctggcgctga
     3421 tggcgcacac catcctcttc ctcaagctct tctcctaccg cgacgtcaac tcatggtgcc
     3481 gcagggccag ggccaaggct gcctctgcag ggaagaaggc cagcagtgct gctgccccgc
     3541 acaccgtgag ctacccggac aatctgacct accgcgatct ctactacttc ctcttcgccc
     3601 ccaccttgtg ctacgagctc aactttcccc gctctccccg catccggaag cgctttctgc
     3661 tgcgacggat ccttgagatg ctgttcttca cccagctcca ggtggggctg atccagcagt
     3721 ggatggtccc caccatccag aactccatga agcccttcaa ggacatggac tactcacgca
     3781 tcatcgagcg cctcctgaag ctggcggtcc ccaatcacct catctggctc atcttcttct
     3841 actggctctt ccactcctgc ctgaatgccg tggctgagct catgcagttt ggagaccggg
     3901 agttctaccg ggactggtgg aactccgagt ctgtcaccta cttctggcag aactggaaca
     3961 tccctgtgca caagtggtgc atcagacact tctacaagcc catgcttcga cggggcagca
     4021 gcaagtggat ggccaggaca ggggtgttcc tggcctcggc cttcttccac gagtacctgg
     4081 tgagcgtccc tctgcgaatg ttccgcctct gggcgttcac gggcatgatg gctcagatcc
     4141 cactggcctg gttcgtgggc cgctttttcc agggcaacta tggcaacgca gctgtgtggc
     4201 tgtcgctcat catcggacag ccaatagccg tcctcatgta cgtccacgac tactacgtgc
     4261 tcaactatga ggccccagcg gcagaggccg aattcgatta caaggatgac gacgataagt
     4321 gaggatcccg cgactctaga taattctacc gggtagggga ggcgcttttc ccaaggcagt
     4381 ctggagcatg cgctttagca gccccgctgg gcacttggcg ctacacaagt ggcctctggc
     4441 ctcgcacaca ttccacatcc accggtaggc gccaaccggc tccgttcttt ggtggcccct
     4501 tcgcgccacc ttctactcct cccctagtca ggaagttccc ccccgccccg cagctcgcgt
     4561 cgtgcaggac gtgacaaatg gaagtagcac gtctcactag tctcgtgcag atggacagca
     4621 ccgctgagca atggaagcgg gtaggccttt ggggcagcgg ccaatagcag ctttgctcct
     4681 tcgctttctg ggctcagagg ctgggaaggg gtgggtccgg gggcgggctc aggggcgggc
     4741 tcaggggcgg ggcgggcgcc cgaaggtcct ccggaggccc ggcattctgc acgcttcaaa
     4801 agcgcacgtc tgccgcgctg ttctcctctt cctcatctcc gggcctttcg acctgcagcc
     4861 caagcttacc atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc
     4921 cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt
     4981 cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt
     5041 cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac
     5101 cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga
     5161 gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg
     5221 gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa
     5281 gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc
     5341 cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac
     5401 cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc
     5461 cggtgcctga ccgcgtctgg aacaatcaac ctctggatta caaaatttgt gaaagattga
     5521 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     5581 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     5641 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     5701 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     5761 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     5821 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc
     5881 tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
     5941 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     6001 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     6061 ccgcctcccc gcctggaatt aattctgcag tcgagaccta gaaaaacatg gagcaatcac
     6121 aagtagcaat acagcagcta ccaatgctga ttgtgcctgg ctagaagcac aagaggagga
     6181 ggaggtgggt tttccagtca cacctcaggt acctttaaga ccaatgactt acaaggcagc
     6241 tgtagatctt agccactttt taaaagaaaa gaggggactg gaagggctaa ttcactccca
     6301 acgaagacaa gatatccttg atctgtggat ctaccacaca caaggctact tccctgatta
     6361 gcagaactac acaccagggc caggggtcag atatccactg acctttggat ggtgctacaa
     6421 gctagtacca gttgagccag ataaggtaga agaggccaat aaaggagaga acaccagctt
     6481 gttacaccct gtgagcctgc atgggatgga tgacccggag agagaagtgt tagagtggag
     6541 gtttgacagc cgcctagcat ttcatcacgt ggcccgagag ctgcatccgg agtacttcaa
     6601 gaactgctga tatcgagctt gctacaaggg actttccgct ggggactttc cagggaggcg
     6661 tggcctgggc gggactgggg agtggcgagc cctcagatcc tgcatataag cagctgcttt
     6721 ttgcctgtac tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
     6781 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
     6841 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
     6901 aaatctctag cagtagtagt tcatgtcatc ttattattca gtatttataa cttgcaaaga
     6961 aatgaatatc agagagtgag aggccttgac attgctagcg ttttaccgtc gacctctagc
     7021 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     7081 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     7141 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     7201 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     7261 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     7321 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     7381 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     7441 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     7501 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     7561 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     7621 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     7681 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     7741 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     7801 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     7861 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     7921 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     7981 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     8041 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     8101 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     8161 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     8221 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     8281 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     8341 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     8401 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     8461 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     8521 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     8581 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     8641 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     8701 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     8761 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     8821 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     8881 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     8941 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     9001 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     9061 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     9121 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     9181 aaagtgccac ctgacgtcga cggatcggga gatcaacttg tttattgcag cttataatgg
     9241 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     9301 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatca actggataac
     9361 tcaagctaac caaaatcatc ccaaacttcc caccccatac cctattacca ctgccaatta
     9421 cctgtggttt catttactct aaacctgtga ttcctctgaa ttattttcat tttaaagaaa
     9481 ttgtatttgt taaatatgta ctacaaactt agtagttttt aaagaaattg tatttgttaa
     9541 atatgtacta caaacttagt agt
//
