LOCUS       V044228                 9983 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044228
VERSION     V044228
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     LTR             234..414
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /label="5' LTR (truncated)"
     misc_feature    458..583
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1076..1309
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1493..1537
                     /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    1804..1920
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2010..2213
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2302..4902
                     /label="E2F8(NM_024680)"
                     /note="E2F8(NM_024680)"
                     /gene="E2F8"
     promoter        4953..5164
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             5177..5445
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5472..6071
                     /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    6078..6666
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6740..6973
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    7045..7166
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      7185..7320
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7358..7374)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7382..7398
                     /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(7406..7436)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7451..7472
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7760..8348)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8519..9379)
                     /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(9380..9484)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9958..9974
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
ORIGIN
        1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
       61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
      121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
      181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc
      241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
      421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct
      481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
      541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
      601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
      661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
      721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
      781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
      841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
      901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg
      961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc
     1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     1381 aatctctgga acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt
     1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaat tcaaaatttt
     1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag
     2221 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct
     2281 agagctagcg aattcgccac catggagaac gaaaaggaaa atctcttttg tgagccacat
     2341 aaaaggggac taatgaaaac acctctgaaa gaatccacca cagcaaatat cgtgttggca
     2401 gagatccagc ctgactttgg ccctttaacc acacctacca agcccaagga aggctctcag
     2461 ggagagccgt ggacaccgac agccaacctg aaaatgctca tcagtgctgt gagccctgag
     2521 atccgcaaca gagatcagaa aaggggtttg tttgacaaca gaagtggatt acctgaggcc
     2581 aaagactgta tacacgaaca cttatctgga gatgaatttg agaaatccca accaagtcga
     2641 aaagagaaaa gtttaggatt attgtgtcat aagttcttag cacgatatcc taattatccc
     2701 aaccctgctg tgaataatga catctgcctt gacgaagtgg cagaggaact taatgttgaa
     2761 cgtcgacgca tttacgatat cgtgaacgtc ctagagagtt tacatatggt gagccgcctc
     2821 gccaaaaaca ggtacacttg gcacgggcga cacaatctca acaaaaccct tggcaccttg
     2881 aagagcatcg gggaggagaa taagtacgcc gagcagatta tgatgatcaa aaagaaagaa
     2941 tatgagcaag agtttgactt tattaagagt tacagtatag aggatcatat catcaaatca
     3001 aacactggcc caaatggaca cccagacatg tgttttgtgg aactccctgg agtggaattt
     3061 cgggcagctt ctgtaaacag ccgcaaagac aagtctttaa gggtaatgag ccagaaattt
     3121 gtgatgctgt ttttggtgtc aacgcctcag atagtaagcc tagaagttgc tgccaagatt
     3181 ttaattgggg aggaccatgt ggaagatttg gataaaagca agtttaaaac aaaaattagg
     3241 aggttgtatg atatagctaa tgttctgagt agcctggatc ttatcaagaa agttcatgtt
     3301 acagaggaaa gaggccgaaa accagctttc aaatggaccg gcccagaaat cagtccaaat
     3361 accagtggct ccagcccagt cattcatttt actccctctg atttggaggt gagacggtct
     3421 tcaaaagaga actgtgccaa aaacctcttt tccacacgtg ggaaaccaaa ctttactcga
     3481 cacccatctc ttatcaaatt ggtaaagagt atagaaagtg atcggagaaa gataaattct
     3541 gcgcccagta gccctatcaa gaccaacaaa gctgagagtt ctcagaattc tgcacccttc
     3601 ccaagtaaaa tggctcagct cgcagctatt tgtaaaatgc agttagaaga gcaatcaagt
     3661 gaatccagac agaaagtgaa agtacagctg gcaagatctg gaccctgcaa accagtagcc
     3721 cctctggacc ccccagtgaa tgctgagatg gagctgacag caccgtccct catccagccc
     3781 ctgggaatgg ttcccctgat ccccagcccc ttgtcatcag cagtgcccct gatcctacct
     3841 caggcccctt caggcccatc ctatgccatc tacctgcagc ccactcaagc ccaccaaagt
     3901 gtgacgccac cccaaggcct gagcccaacg gtgtgcacca cccactcttc taaagctact
     3961 ggctcaaaag actccacaga tgccaccact gagaaggcag ccaatgatac ctcaaaggcc
     4021 agtgcctcta ccaggcctgg aagcttgctg ccagcaccag agaggcaagg ggcaaagagc
     4081 cgaaccaggg agccagctgg agaaagaggc tcaaagaggg caagcatgct cgaggacagt
     4141 ggttccaaaa agaaatttaa agaggaccta aaaggacttg aaaatgtctc cgcaaccttg
     4201 ttcccatcag gatacctaat ccctctcacg cagtgctcat ccctgggggc agagtccatt
     4261 ttgtctggta aagaaaactc aagtgctctt tccccaaacc acaggattta cagctcccca
     4321 attgcaggtg ttattccagt gacatcatct gaactcactg ctgttaattt tccctctttt
     4381 catgtaacac cgttgaagct aatggtctca ccaacttccg tggcagccgt acctgtcggg
     4441 aacagcccgg ctctcgcttc aagccaccct gttcccatcc agaacccaag ctcagccatt
     4501 gtaaacttca ccctgcagca cctgggactc atctcaccca atgtgcagtt gtctgccagc
     4561 cctgggtctg gaatcgttcc tgtgtctcca agaatagagt ctgttaatgt cgcaccagaa
     4621 aatgcaggca ctcagcaagg aagggccacc aactatgact caccagtccc aggccagagc
     4681 cagccaaatg gacaatcagt tgctgtgaca ggggcacaac agcctgttcc tgtgacaccc
     4741 aaagggtcac aattagtggc cgaaagtttc ttccgtaccc caggtggacc caccaagcca
     4801 accagctcat cctgcatgga ttttgagggt gctaataaaa cctccttagg aactctcttt
     4861 gtcccacagc gaaaactgga agtctcaaca gaggatgtcc attaaggatc cgcggccgcg
     4921 aaggatctgc gatcgctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc
     4981 cgagaagttg gggggagggg tcggcaattg aacgggtgcc tagagaaggt ggcgcggggt
     5041 aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc
     5101 gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac
     5161 acagctgaag cttcgagggg ctcgcatctc tccttcacgc gcccgccgcc ctacctgagg
     5221 ccgccatcca cgccggttga gtcgcgttct gccgcctccc gcctgtggtg cctcctgaac
     5281 tgcgtccgcc gtctaggtaa gtttaaagct caggtcgaga ccgggccttt gtccggcgct
     5341 cccttggagc ctacctagac tcagccggct ctccacgctt tgcctgaccc tgcttgctca
     5401 actctacgtc tttgtttcgt tttctgttct gcgccgttac agatccaagc tgtgaccggc
     5461 gcctacgcta gatgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc
     5521 cccgggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg
     5581 tcgacccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg
     5641 tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga
     5701 ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg
     5761 agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc
     5821 ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca
     5881 agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc
     5941 ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca
     6001 ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc
     6061 ccggtgcctg agtcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     6121 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     6181 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     6241 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     6301 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     6361 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     6421 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt
     6481 cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct
     6541 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     6601 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     6661 ccccgcctgg tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt
     6721 ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaaaata agatctgctt
     6781 tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa
     6841 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
     6901 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
     6961 aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag
     7021 aaatgaatat cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa
     7081 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     7141 ttgtccaaac tcatcaatgt atcttatcat gtctggctct agctatcccg cccctaactc
     7201 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     7261 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     7321 tagacttttg cagagacggc ccaaattcgt aatcatggtc atagctgttt cctgtgtgaa
     7381 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     7441 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     7501 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     7561 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     7621 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     7681 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     7741 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     7801 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     7861 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     7921 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     7981 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     8041 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     8101 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     8161 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     8221 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     8281 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     8341 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     8401 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     8461 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     8521 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     8581 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     8641 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     8701 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     8761 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     8821 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     8881 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     8941 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     9001 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     9061 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     9121 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     9181 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     9241 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     9301 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     9361 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     9421 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     9481 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     9541 taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg
     9601 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     9661 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
     9721 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac
     9781 cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca
     9841 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
     9901 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
     9961 aaacgacggc cagtgccaag ctg
//
