LOCUS       V031598                 9639 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031598
VERSION     V031598
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             2830..2856
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2863..4395
                     /label="APLF(NM_173545)"
                     /note="APLF(NM_173545)"
                     /gene="APLF"
     promoter        4427..4926
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             4947..5546
                     /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    5560..6148
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6355..6988
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7117..7133)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7141..7157
                     /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(7165..7195)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7210..7231
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7519..8107)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8278..9138)
                     /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(9139..9243)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9291..9425
                     /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 gccaccatgt acccatacga cgtaccagat tacgctgaat tcatgtccgg gggcttcgag
     2881 ctgcagccgc gggacggcgg tccccgggtg gccctggcgc ccggggagac ggtgatcggc
     2941 cgcgggccgc tgctgggaat aacagacaag agagtatcca gaagacatgc cattcttgag
     3001 gtggcaggtg gtcagctgcg aatcaaaccg atacacacaa atccatgttt ttaccagtct
     3061 tcagagaaga gtcagctctt accattgaag ccaaatctat ggtgctattt gaatcctgga
     3121 gacagctttt ctttgttagt tgacaaatac attttccgca ttctctctat accctctgaa
     3181 gtggaaatgc aatgtacctt aagaaacagt caagtgcttg atgaagataa tatattgaat
     3241 gaaacaccaa aatcccccgt gattaattta cctcatgaga ctactggtgc ctcacaactg
     3301 gaaggaagca cagaaatagc caagacccag atgactccca caaatagtgt gtctttccta
     3361 ggtgaaaata gagactgcaa taagcagcag ccaatccttg ccgagaggaa aagaatcctt
     3421 ccaacttgga tgttagcaga acatttaagt gatcaaaacc tttcagtacc agcaatcagt
     3481 ggaggtaatg taatccaggg aagtggaaaa gaagaaatct gcaaagataa atcccagcta
     3541 aacacaaccc agcaaggaag aaggcaatta atttcatcag gaagttcaga aaatacatca
     3601 gcagaacaag acacaggaga agagtgcaaa aatactgatc aggaagagtc taccatttca
     3661 tccaaggaaa tgccacaatc attttctgca atcacattaa gtaacacaga gatgaataat
     3721 attaagacta atgcacagag aaacaaactt ccaatagagg aacttggtaa agtttctaaa
     3781 cataaaattg ccactaaaag aacaccacat aaagaagatg aggcaatgag ctgttctgaa
     3841 aattgttcga gtgcccaggg cgactcactt caggatgagt ctcaagggtc tcattctgag
     3901 tccagctcta atccctccaa tcctgaaact ttgcatgcaa aggcaactga ttcagttcta
     3961 caaggttctg aaggaaacaa ggtcaagagg acatcctgca tgtatggggc aaactgctat
     4021 aggaagaatc ctgttcattt tcaacatttt agccatcctg gtgatagtga ttatggaggt
     4081 gtacaaatcg tgggccaaga tgagactgat gaccggcctg aatgtcccta tggaccatcc
     4141 tgttatagga agaatcccca gcacaagata gaatatagac ataatacgct tccagtgaga
     4201 aatgttttag atgaagataa tgataatgtt gggcaaccca atgagtatga cctgaacgac
     4261 agctttctag atgatgagga agaagactat gagccaacag atgaagattc tgactgggaa
     4321 ccaggaaagg aagatgaaga gaaggaagat gtggaagagc ttttgaaaga agcaaaaagg
     4381 tttatgaaaa gaaaataggg atcccgcgac tctagataat tctaccgggt aggggaggcg
     4441 cttttcccaa ggcagtctgg agcatgcgct ttagcagccc cgctgggcac ttggcgctac
     4501 acaagtggcc tctggcctcg cacacattcc acatccaccg gtaggcgcca accggctccg
     4561 ttctttggtg gccccttcgc gccaccttct actcctcccc tagtcaggaa gttccccccc
     4621 gccccgcagc tcgcgtcgtg caggacgtga caaatggaag tagcacgtct cactagtctc
     4681 gtgcagatgg acagcaccgc tgagcaatgg aagcgggtag gcctttgggg cagcggccaa
     4741 tagcagcttt gctccttcgc tttctgggct cagaggctgg gaaggggtgg gtccgggggc
     4801 gggctcaggg gcgggctcag gggcggggcg ggcgcccgaa ggtcctccgg aggcccggca
     4861 ttctgcacgc ttcaaaagcg cacgtctgcc gcgctgttct cctcttcctc atctccgggc
     4921 ctttcgacct gcagcccaag cttaccatga ccgagtacaa gcccacggtg cgcctcgcca
     4981 cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg
     5041 ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac
     5101 tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg
     5161 cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg
     5221 gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc
     5281 tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc
     5341 ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg
     5401 agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg
     5461 agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt
     5521 gcatgacccg caagcccggt gcctgaccgc gtctggaaca atcaacctct ggattacaaa
     5581 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     5641 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     5701 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     5761 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     5821 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     5881 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     5941 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt
     6001 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     6061 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     6121 cggatctccc tttgggccgc ctccccgcct ggaattaatt ctgcagtcga gacctagaaa
     6181 aacatggagc aatcacaagt agcaatacag cagctaccaa tgctgattgt gcctggctag
     6241 aagcacaaga ggaggaggag gtgggttttc cagtcacacc tcaggtacct ttaagaccaa
     6301 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagagg ggactggaag
     6361 ggctaattca ctcccaacga agacaagata tccttgatct gtggatctac cacacacaag
     6421 gctacttccc tgattagcag aactacacac cagggccagg ggtcagatat ccactgacct
     6481 ttggatggtg ctacaagcta gtaccagttg agccagataa ggtagaagag gccaataaag
     6541 gagagaacac cagcttgtta caccctgtga gcctgcatgg gatggatgac ccggagagag
     6601 aagtgttaga gtggaggttt gacagccgcc tagcatttca tcacgtggcc cgagagctgc
     6661 atccggagta cttcaagaac tgctgatatc gagcttgcta caagggactt tccgctgggg
     6721 actttccagg gaggcgtggc ctgggcggga ctggggagtg gcgagccctc agatcctgca
     6781 tataagcagc tgctttttgc ctgtactggg tctctctggt tagaccagat ctgagcctgg
     6841 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
     6901 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
     6961 ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat
     7021 ttataacttg caaagaaatg aatatcagag agtgagaggc cttgacattg ctagcgtttt
     7081 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     7141 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     7201 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     7261 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     7321 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     7381 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     7441 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     7501 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     7561 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     7621 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     7681 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     7741 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     7801 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     7861 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     7921 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     7981 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     8041 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     8101 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     8161 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     8221 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     8281 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     8341 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     8401 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     8461 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     8521 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     8581 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     8641 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     8701 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     8761 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     8821 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     8881 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     8941 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     9001 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     9061 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     9121 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     9181 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     9241 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc aacttgttta
     9301 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     9361 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     9421 ggatcaactg gataactcaa gctaaccaaa atcatcccaa acttcccacc ccatacccta
     9481 ttaccactgc caattacctg tggtttcatt tactctaaac ctgtgattcc tctgaattat
     9541 tttcatttta aagaaattgt atttgttaaa tatgtactac aaacttagta gtttttaaag
     9601 aaattgtatt tgttaaatat gtactacaaa cttagtagt
//
