LOCUS       V036607                 8997 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036607
VERSION     V036607
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             2827..3759
                     /label="Spp1(NM_001204201)"
                     /note="Spp1(NM_001204201)"
                     /gene="Spp1"
     promoter        3785..4284
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             4305..4904
                     /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    4918..5506
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5713..6346
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(6475..6491)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6499..6515
                     /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(6523..6553)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6568..6589
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6877..7465)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7636..8496)
                     /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(8497..8601)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    8649..8783
                     /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 gccaccatga ggctgcagtt ctcctggctg aattctgagg gactaactac gaccatgaga
     2881 ttggcagtga tttgcttttg cctgtttggc attgcctcct ccctcccggt gaaagtgact
     2941 gattctggca gctcagagga gaagaagctt tacagcctgc acccagatcc tatagccaca
     3001 tggctggtgc ctgacccatc tcagaagcag aatctccttg cgccacagaa tgctgtgtcc
     3061 tctgaagaaa aggatgactt taagcaagaa actcttccaa gcaattccaa tgaaagccat
     3121 gaccacatgg acgacgatga tgacgatgat gatgacgatg gagaccatgc agagagcgag
     3181 gattctgtgg actcggatga atctgacgaa tctcaccatt cggatgagtc tgatgagacc
     3241 gtcactgcta gtacacaagc agacactttc actccaatcg tccctacagt cgatgtcccc
     3301 aacggccgag gtgatagctt ggcttatgga ctgaggtcaa agtctaggag tttccaggtt
     3361 tctgatgaac agtatcctga tgccacagat gaggacctca cctctcacat gaagagcggt
     3421 gagtctaagg agtccctcga tgtcatccct gttgcccagc ttctgagcat gccctctgat
     3481 caggacaaca acggaaaggg cagccatgag tcaagtcagc tggatgaacc aagtctggaa
     3541 acacacagac ttgagcattc caaagagagc caggagagtg ccgatcagtc ggatgtgatc
     3601 gatagtcaag caagttccaa agccagcctg gaacatcaga gccacaagtt tcacagccac
     3661 aaggacaagc tagtcctaga ccctaagagt aaggaagatg ataggtatct gaaattccga
     3721 atttctcatg aattagagag ttcatcttct gaggtcaact aaggatcctc tagataattc
     3781 taccgggtag gggaggcgct tttcccaagg cagtctggag catgcgcttt agcagccccg
     3841 ctgggcactt ggcgctacac aagtggcctc tggcctcgca cacattccac atccaccggt
     3901 aggcgccaac cggctccgtt ctttggtggc cccttcgcgc caccttctac tcctccccta
     3961 gtcaggaagt tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca aatggaagta
     4021 gcacgtctca ctagtctcgt gcagatggac agcaccgctg agcaatggaa gcgggtaggc
     4081 ctttggggca gcggccaata gcagctttgc tccttcgctt tctgggctca gaggctggga
     4141 aggggtgggt ccgggggcgg gctcaggggc gggctcaggg gcggggcggg cgcccgaagg
     4201 tcctccggag gcccggcatt ctgcacgctt caaaagcgca cgtctgccgc gctgttctcc
     4261 tcttcctcat ctccgggcct ttcgacctgc agcccaagct taccatgacc gagtacaagc
     4321 ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg
     4381 cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg
     4441 tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg
     4501 tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg
     4561 cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc
     4621 agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg
     4681 ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc
     4741 ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc
     4801 gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg
     4861 aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgaccgcgt ctggaacaat
     4921 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     4981 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     5041 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     5101 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     5161 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     5221 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     5281 ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc
     5341 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     5401 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     5461 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg aattaattct
     5521 gcagtcgaga cctagaaaaa catggagcaa tcacaagtag caatacagca gctaccaatg
     5581 ctgattgtgc ctggctagaa gcacaagagg aggaggaggt gggttttcca gtcacacctc
     5641 aggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag
     5701 aaaagagggg actggaaggg ctaattcact cccaacgaag acaagatatc cttgatctgt
     5761 ggatctacca cacacaaggc tacttccctg attagcagaa ctacacacca gggccagggg
     5821 tcagatatcc actgaccttt ggatggtgct acaagctagt accagttgag ccagataagg
     5881 tagaagaggc caataaagga gagaacacca gcttgttaca ccctgtgagc ctgcatggga
     5941 tggatgaccc ggagagagaa gtgttagagt ggaggtttga cagccgccta gcatttcatc
     6001 acgtggcccg agagctgcat ccggagtact tcaagaactg ctgatatcga gcttgctaca
     6061 agggactttc cgctggggac tttccaggga ggcgtggcct gggcgggact ggggagtggc
     6121 gagccctcag atcctgcata taagcagctg ctttttgcct gtactgggtc tctctggtta
     6181 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     6241 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     6301 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt
     6361 catcttatta ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggcct
     6421 tgacattgct agcgttttac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     6481 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     6541 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     6601 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     6661 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     6721 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     6781 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     6841 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     6901 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     6961 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     7021 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     7081 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     7141 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     7201 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     7261 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     7321 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     7381 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     7441 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     7501 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     7561 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     7621 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     7681 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     7741 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     7801 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     7861 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     7921 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     7981 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     8041 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     8101 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     8161 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     8221 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     8281 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     8341 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     8401 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     8461 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     8521 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     8581 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
     8641 gggagatcaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     8701 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     8761 atgtatctta tcatgtctgg atcaactgga taactcaagc taaccaaaat catcccaaac
     8821 ttcccacccc ataccctatt accactgcca attacctgtg gtttcattta ctctaaacct
     8881 gtgattcctc tgaattattt tcattttaaa gaaattgtat ttgttaaata tgtactacaa
     8941 acttagtagt ttttaaagaa attgtatttg ttaaatatgt actacaaact tagtagt
//
