LOCUS       V039778                10306 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V039778
VERSION     V039778
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             2843..5002
                     /label="Acsl3(NM_028817)"
                     /note="Acsl3(NM_028817)"
                     /gene="Acsl3"
     CDS             5009..5074
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        5094..5593
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5614..6213
                     /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    6227..6815
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             7022..7655
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7784..7800)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7808..7824
                     /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(7832..7862)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7877..7898
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8186..8774)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8945..9805)
                     /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(9806..9910)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9958..10092
                     /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 ctcaagcttc gaattcgcca ccatgaataa ccacgtatct tcaacaccat caaccatgaa
     2881 gctaaaacaa accatcaacc ccatactttt atatttcata cattttataa tatcactcta
     2941 tactatttta acatacatcc cattttattt tttgtgtgag tcaaaacaag agaaaccaaa
     3001 ccaaattaaa gcaaaacctg tcagttccaa accggactct gcatacagat ctatcaacag
     3061 tgtggatggc ttggcttcag tgctgtatcc tggctgcgat acacttgata aagtctttat
     3121 gtatgcaaaa aacaaattta aaaacaaaag actattggga acacgtgaaa ttttaaatga
     3181 ggaagatgaa atacagccaa atggaaaaat ttttaaaaag gttattctgg ggcactataa
     3241 ttggctttcc tatgaagatg tcttcatccg agcccttgac tttggaaatg ggttacaaat
     3301 gttgggccag aaaccaaagg ccaacatcgc catcttctgt gagaccaggg ctgagtggat
     3361 gatcgctgca caggcgtgtt ttatgtataa cttccagctt gttacactgt atgctactct
     3421 gggaggtcca gccattgttc atggactgaa tgagacagag gtgaccaaca tcattactag
     3481 caaagaactc ttgcaaacaa agctgaagga tatagtctct ttggtcccac gtctgcggca
     3541 tatcattact gttgatggga agcctccaac ctggtctgag ttccccaaag gtgtcattgt
     3601 acacaccatg gctgcagtgc aggctctagg agtgaaggcc aacgtggaaa agaaagctca
     3661 cagcaaacca ctgccctcag atattgcagt aatcatgtac acaagtgggt ccacaggaat
     3721 tccaaaggga gtcatgatct cacacagcaa catcattgct tctataacgg ggatggcgag
     3781 aaggattcca agactgggag aggaagatgt gtatattggc tacttgcccc tggcacatgt
     3841 tctagaatta agtgctgagc ttgtgtgtct ttctcatgga tgccgaatcg gctactcttc
     3901 accacagaca ttagcagatc agtcttcaaa aataaaaaaa ggaagcaaag gagacacatc
     3961 cgtgctgaag ccaacactga tggcagctgt tccggaaatc atggatcgga tctacaaaaa
     4021 tgtcatgaat aaagtgaatg aaatgagtgc ttttcaacga aacttgttta ttttggcata
     4081 taattataag atggaacaga tttcaaaagg gtgtagtact ccactgtgtg accgctttgt
     4141 tttccggaat gtccgaaggc tgctgggtgg aaatattcgc cttttactgt gtggtggtgc
     4201 tccgctttct gcaacaacgc agcgattcat gaatatctgt ttctgctgtc ccgttggtca
     4261 ggggtatgga ctcacagaat ctactggggc tggaacaatt acagaagtgt gggactacaa
     4321 taccggcaga gtgggagcac cattagtttg ctgtgaaatc aaattaaaga actgggagga
     4381 aggtggctat tttaatactg acaaaccaca tcccagaggt gaaattctta ttggtggcca
     4441 aaatgtgaca atggggtact acaaaaatga agcaaaaaca aagacagatt tctttgaaga
     4501 tgaaaatgga cagcggtggc tgtgcactgg agatattgga gagtttgacc ctgacggctg
     4561 tctgaagatc attgaccgta aaaaggacct tgtgaaacta caggcaggag agtacgtttc
     4621 tctcgggaaa gtagaggcag ctttgaagaa cctcccactg atagataaca tttgtgcgta
     4681 tgcaaacagc taccattctt acgtaattgg gtttgttgtg ccaaatcaaa aggaacttac
     4741 agagctagct agaacaaaag gatttaaagg aacttgggaa gagctgtgta acagcagcga
     4801 gatggaaaat gaggtcctta aagtgctttc tgaagctgct atttcagcaa gtctggaaaa
     4861 gtttgaaatt ccactaaaaa tacgtttgag ccctgaccca tggactcccg aaactggtct
     4921 ggtgacagat gccttcaagt tgaaacgtaa agagcttaaa acacactacc aggcagacat
     4981 tgagcggatg tatggaagaa aaggatccga ctacaaagac catgacggtg attataaaga
     5041 tcatgacatc gactacaagg atgacgatga caagtagtct agataattct accgggtagg
     5101 ggaggcgctt ttcccaaggc agtctggagc atgcgcttta gcagccccgc tgggcacttg
     5161 gcgctacaca agtggcctct ggcctcgcac acattccaca tccaccggta ggcgccaacc
     5221 ggctccgttc tttggtggcc ccttcgcgcc accttctact cctcccctag tcaggaagtt
     5281 cccccccgcc ccgcagctcg cgtcgtgcag gacgtgacaa atggaagtag cacgtctcac
     5341 tagtctcgtg cagatggaca gcaccgctga gcaatggaag cgggtaggcc tttggggcag
     5401 cggccaatag cagctttgct ccttcgcttt ctgggctcag aggctgggaa ggggtgggtc
     5461 cgggggcggg ctcaggggcg ggctcagggg cggggcgggc gcccgaaggt cctccggagg
     5521 cccggcattc tgcacgcttc aaaagcgcac gtctgccgcg ctgttctcct cttcctcatc
     5581 tccgggcctt tcgacctgca gcccaagctt accatgaccg agtacaagcc cacggtgcgc
     5641 ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac
     5701 taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg
     5761 caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac
     5821 ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc
     5881 gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg
     5941 gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc
     6001 gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag
     6061 gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc
     6121 ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc
     6181 acctggtgca tgacccgcaa gcccggtgcc tgaccgcgtc tggaacaatc aacctctgga
     6241 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     6301 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     6361 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     6421 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     6481 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     6541 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     6601 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac
     6661 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     6721 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     6781 gacgagtcgg atctcccttt gggccgcctc cccgcctgga attaattctg cagtcgagac
     6841 ctagaaaaac atggagcaat cacaagtagc aatacagcag ctaccaatgc tgattgtgcc
     6901 tggctagaag cacaagagga ggaggaggtg ggttttccag tcacacctca ggtaccttta
     6961 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagagggga
     7021 ctggaagggc taattcactc ccaacgaaga caagatatcc ttgatctgtg gatctaccac
     7081 acacaaggct acttccctga ttagcagaac tacacaccag ggccaggggt cagatatcca
     7141 ctgacctttg gatggtgcta caagctagta ccagttgagc cagataaggt agaagaggcc
     7201 aataaaggag agaacaccag cttgttacac cctgtgagcc tgcatgggat ggatgacccg
     7261 gagagagaag tgttagagtg gaggtttgac agccgcctag catttcatca cgtggcccga
     7321 gagctgcatc cggagtactt caagaactgc tgatatcgag cttgctacaa gggactttcc
     7381 gctggggact ttccagggag gcgtggcctg ggcgggactg gggagtggcg agccctcaga
     7441 tcctgcatat aagcagctgc tttttgcctg tactgggtct ctctggttag accagatctg
     7501 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
     7561 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
     7621 cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat
     7681 tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggcctt gacattgcta
     7741 gcgttttacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     7801 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     7861 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     7921 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     7981 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     8041 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     8101 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     8161 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     8221 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     8281 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     8341 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     8401 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     8461 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     8521 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     8581 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     8641 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     8701 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     8761 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     8821 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     8881 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     8941 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     9001 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     9061 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     9121 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     9181 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     9241 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     9301 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     9361 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     9421 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     9481 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     9541 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     9601 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     9661 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     9721 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     9781 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     9841 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     9901 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatcaac
     9961 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
    10021 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
    10081 catgtctgga tcaactggat aactcaagct aaccaaaatc atcccaaact tcccacccca
    10141 taccctatta ccactgccaa ttacctgtgg tttcatttac tctaaacctg tgattcctct
    10201 gaattatttt cattttaaag aaattgtatt tgttaaatat gtactacaaa cttagtagtt
    10261 tttaaagaaa ttgtatttgt taaatatgta ctacaaactt agtagt
//
