LOCUS       pLVX-Puro-CBS(human)    9739 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043941
VERSION     V043941
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..4495
                     /label="CBS(NM_000071)"
                     /note="CBS(NM_000071)"
                     /gene="CBS"
     promoter        4527..5026
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5047..5646
                     /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    5660..6248
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6455..7088
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7217..7233)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7241..7257
                     /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(7265..7295)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7310..7331
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7619..8207)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8378..9238)
                     /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(9239..9343)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9391..9525
                     /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 ccatgccttc tgagaccccc caggcagaag tggggcccac
     2881 aggctgcccc caccgctcag ggccacactc ggcgaagggg agcctggaga aggggtcccc
     2941 agaggataag gaagccaagg agcccctgtg gatccggccc gatgctccga gcaggtgcac
     3001 ctggcagctg ggccggcctg cctccgagtc cccacatcac cacactgccc cggcaaaatc
     3061 tccaaaaatc ttgccagata ttctgaagaa aatcggggac acccctatgg tcagaatcaa
     3121 caagattggg aagaagttcg gcctgaagtg tgagctcttg gccaagtgtg agttcttcaa
     3181 cgcgggcggg agcgtgaagg accgcatcag cctgcggatg attgaggatg ctgagcgcga
     3241 cgggacgctg aagcccgggg acacgattat cgagccgaca tccgggaaca ccgggatcgg
     3301 gctggccctg gctgcggcag tgaggggcta tcgctgcatc atcgtgatgc cagagaagat
     3361 gagctccgag aaggtggacg tgctgcgggc actgggggct gagattgtga ggacgcccac
     3421 caatgccagg ttcgactccc cggagtcaca cgtgggggtg gcctggcggc tgaagaacga
     3481 aatccccaat tctcacatcc tagaccagta ccgcaacgcc agcaaccccc tggctcacta
     3541 cgacaccacc gctgatgaga tcctgcagca gtgtgatggg aagctggaca tgctggtggc
     3601 ttcagtgggc acgggcggca ccatcacggg cattgccagg aagctgaagg agaagtgtcc
     3661 tggatgcagg atcattgggg tggatcccga agggtccatc ctcgcagagc cggaggagct
     3721 gaaccagacg gagcagacaa cctacgaggt ggaagggatc ggctacgact tcatccccac
     3781 ggtgctggac aggacggtgg tggacaagtg gttcaagagc aacgatgagg aggcgttcac
     3841 ctttgcccgc atgctgatcg cgcaagaggg gctgctgtgc ggtggcagtg ctggcagcac
     3901 ggtggcggtg gccgtgaagg ccgcgcagga gctgcaggag ggccagcgct gcgtggtcat
     3961 tctgcccgac tcagtgcgga actacatgac caagttcctg agcgacaggt ggatgctgca
     4021 gaagggcttt ctgaaggagg aggacctcac ggagaagaag ccctggtggt ggcacctccg
     4081 tgttcaggag ctgggcctgt cagccccgct gaccgtgctc ccgaccatca cctgtgggca
     4141 caccatcgag atcctccggg agaagggctt cgaccaggcg cccgtggtgg atgaggcggg
     4201 ggtaatcctg ggaatggtga cgcttgggaa catgctctcg tccctgcttg ccgggaaggt
     4261 gcagccgtca gaccaagttg gcaaagtcat ctacaagcag ttcaaacaga tccgcctcac
     4321 ggacacgctg ggcaggctct cgcacatcct ggagatggac cacttcgccc tggtggtgca
     4381 cgagcagatc cagtaccaca gcaccgggaa gtccagtcag cggcagatgg tgttcggggt
     4441 ggtcaccgcc attgacttgc tgaacttcgt ggccgcccag gagcgggacc agaagtgagg
     4501 atcccgcgac tctagataat tctaccgggt aggggaggcg cttttcccaa ggcagtctgg
     4561 agcatgcgct ttagcagccc cgctgggcac ttggcgctac acaagtggcc tctggcctcg
     4621 cacacattcc acatccaccg gtaggcgcca accggctccg ttctttggtg gccccttcgc
     4681 gccaccttct actcctcccc tagtcaggaa gttccccccc gccccgcagc tcgcgtcgtg
     4741 caggacgtga caaatggaag tagcacgtct cactagtctc gtgcagatgg acagcaccgc
     4801 tgagcaatgg aagcgggtag gcctttgggg cagcggccaa tagcagcttt gctccttcgc
     4861 tttctgggct cagaggctgg gaaggggtgg gtccgggggc gggctcaggg gcgggctcag
     4921 gggcggggcg ggcgcccgaa ggtcctccgg aggcccggca ttctgcacgc ttcaaaagcg
     4981 cacgtctgcc gcgctgttct cctcttcctc atctccgggc ctttcgacct gcagcccaag
     5041 cttaccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg
     5101 gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat
     5161 ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg
     5221 ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg
     5281 ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg
     5341 agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc
     5401 aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt
     5461 ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc
     5521 ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc
     5581 accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt
     5641 gcctgaccgc gtctggaaca atcaacctct ggattacaaa atttgtgaaa gattgactgg
     5701 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     5761 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     5821 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     5881 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     5941 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     6001 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac
     6061 gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     6121 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     6181 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     6241 ctccccgcct ggaattaatt ctgcagtcga gacctagaaa aacatggagc aatcacaagt
     6301 agcaatacag cagctaccaa tgctgattgt gcctggctag aagcacaaga ggaggaggag
     6361 gtgggttttc cagtcacacc tcaggtacct ttaagaccaa tgacttacaa ggcagctgta
     6421 gatcttagcc actttttaaa agaaaagagg ggactggaag ggctaattca ctcccaacga
     6481 agacaagata tccttgatct gtggatctac cacacacaag gctacttccc tgattagcag
     6541 aactacacac cagggccagg ggtcagatat ccactgacct ttggatggtg ctacaagcta
     6601 gtaccagttg agccagataa ggtagaagag gccaataaag gagagaacac cagcttgtta
     6661 caccctgtga gcctgcatgg gatggatgac ccggagagag aagtgttaga gtggaggttt
     6721 gacagccgcc tagcatttca tcacgtggcc cgagagctgc atccggagta cttcaagaac
     6781 tgctgatatc gagcttgcta caagggactt tccgctgggg actttccagg gaggcgtggc
     6841 ctgggcggga ctggggagtg gcgagccctc agatcctgca tataagcagc tgctttttgc
     6901 ctgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
     6961 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
     7021 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat
     7081 ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg
     7141 aatatcagag agtgagaggc cttgacattg ctagcgtttt accgtcgacc tctagctaga
     7201 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     7261 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     7321 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     7381 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     7441 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     7501 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     7561 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     7621 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     7681 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     7741 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     7801 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     7861 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     7921 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     7981 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     8041 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     8101 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     8161 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     8221 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     8281 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     8341 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     8401 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     8461 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     8521 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     8581 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     8641 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     8701 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     8761 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     8821 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     8881 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     8941 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     9001 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     9061 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     9121 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     9181 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     9241 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     9301 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     9361 tgccacctga cgtcgacgga tcgggagatc aacttgttta ttgcagctta taatggttac
     9421 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     9481 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatcaactg gataactcaa
     9541 gctaaccaaa atcatcccaa acttcccacc ccatacccta ttaccactgc caattacctg
     9601 tggtttcatt tactctaaac ctgtgattcc tctgaattat tttcatttta aagaaattgt
     9661 atttgttaaa tatgtactac aaacttagta gtttttaaag aaattgtatt tgttaaatat
     9721 gtactacaaa cttagtagt
//
