LOCUS       V040437                 8730 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V040437
VERSION     V040437
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..3612
                     /label="E2F1(NM_005225)"
                     /note="E2F1(NM_005225)"
                     /gene="E2F1"
     CDS             3619..3648
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     promoter        3692..3903
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3916..4184
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4219..4818
                     /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    4825..5413
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5487..5720
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5792..5913
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      5932..6067
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6105..6121)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6129..6145
                     /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(6153..6183)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6198..6219
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6507..7095)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7266..8126)
                     /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(8127..8231)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8705..8721
                     /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 catggccttg gccggggccc ctgcgggcgg cccatgcgcg
     2341 ccggcgctgg aggccctgct cggggccggc gcgctgcggc tgctcgactc ctcgcagatc
     2401 gtcatcatct ccgccgcgca ggacgccagc gccccgccgg ctcccaccgg ccccgcggcg
     2461 cccgccgccg gcccctgcga ccctgacctg ctgctcttcg ccacaccgca ggcgccccgg
     2521 cccacaccca gtgcgccgcg gcccgcgctc ggccgcccgc cggtgaagcg gaggctggac
     2581 ctggaaactg accatcagta cctggccgag agcagtgggc cagctcgggg cagaggccgc
     2641 catccaggaa aaggtgtgaa atccccgggg gagaagtcac gctatgagac ctcactgaat
     2701 ctgaccacca agcgcttcct ggagctgctg agccactcgg ctgacggtgt cgtcgacctg
     2761 aactgggctg ccgaggtgct gaaggtgcag aagcggcgca tctatgacat caccaacgtc
     2821 cttgagggca tccagctcat tgccaagaag tccaagaacc acatccagtg gctgggcagc
     2881 cacaccacag tgggcgtcgg cggacggctt gaggggttga cccaggacct ccgacagctg
     2941 caggagagcg agcagcagct ggaccacctg atgaatatct gtactacgca gctgcgcctg
     3001 ctctccgagg acactgacag ccagcgcctg gcctacgtga cgtgtcagga ccttcgtagc
     3061 attgcagacc ctgcagagca gatggttatg gtgatcaaag cccctcctga gacccagctc
     3121 caagccgtgg actcttcgga gaactttcag atctccctta agagcaaaca aggcccgatc
     3181 gatgttttcc tgtgccctga ggagaccgta ggtgggatca gccctgggaa gaccccatcc
     3241 caggaggtca cttctgagga ggagaacagg gccactgact ctgccaccat agtgtcacca
     3301 ccaccatcat ctcccccctc atccctcacc acagatccca gccagtctct actcagcctg
     3361 gagcaagaac cgctgttgtc ccggatgggc agcctgcggg ctcccgtgga cgaggaccgc
     3421 ctgtccccgc tggtggcggc cgactcgctc ctggagcatg tgcgggagga cttctccggc
     3481 ctcctccctg aggagttcat cagcctttcc ccaccccacg aggccctcga ctaccacttc
     3541 ggcctcgagg agggcgaggg catcagagac ctcttcgact gtgactttgg ggacctcacc
     3601 cccctggatt tcctcgagga acaaaaacta atatcagagg aagatttgtg agcggccgca
     3661 aggatctgcg atcgctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc
     3721 gagaagttgg ggggaggggt cggcaattga acgggtgcct agagaaggtg gcgcggggta
     3781 aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg
     3841 tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca
     3901 cagctgaagc ttcgaggggc tcgcatctct ccttcacgcg cccgccgccc tacctgaggc
     3961 cgccatccac gccggttgag tcgcgttctg ccgcctcccg cctgtggtgc ctcctgaact
     4021 gcgtccgccg tctaggtaag tttaaagctc aggtcgagac cgggcctttg tccggcgctc
     4081 ccttggagcc tacctagact cagccggctc tccacgcttt gcctgaccct gcttgctcaa
     4141 ctctacgtct ttgtttcgtt ttctgttctg cgccgttaca gatccaagct gtgaccggcg
     4201 cctaccccgg gcggccctat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac
     4261 gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc
     4321 cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc
     4381 acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg
     4441 gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc
     4501 atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg
     4561 ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac
     4621 cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc
     4681 ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc
     4741 ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc
     4801 cgcaagcccg gtgcctgagt cgacaatcaa cctctggatt acaaaatttg tgaaagattg
     4861 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     4921 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     4981 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     5041 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     5101 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     5161 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
     5221 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     5281 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     5341 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     5401 gccgcctccc cgcctggtac ctttaagacc aatgacttac aaggcagctg tagatcttag
     5461 ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac gaaaataaga
     5521 tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc
     5581 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
     5641 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
     5701 agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt atttataact
     5761 tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt ataatggtta
     5821 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     5881 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc tatcccgccc
     5941 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     6001 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     6061 ggaggcctag acttttgcag agacggccca aattcgtaat catggtcata gctgtttcct
     6121 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     6181 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     6241 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     6301 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     6361 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     6421 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     6481 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     6541 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     6601 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     6661 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     6721 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     6781 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     6841 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     6901 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     6961 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     7021 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     7081 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     7141 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     7201 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     7261 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     7321 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     7381 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     7441 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     7501 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     7561 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     7621 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     7681 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     7741 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     7801 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     7861 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     7921 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     7981 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     8041 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     8101 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     8161 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     8221 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     8281 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt
     8341 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     8401 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     8461 ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt
     8521 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc gccattcagg
     8581 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg
     8641 aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
     8701 cgttgtaaaa cgacggccag tgccaagctg
//
