LOCUS       V019724                 9313 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019724
VERSION     V019724
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             2294..3484
                     /label="DNAJA1(NM_001539)"
                     /note="DNAJA1(NM_001539)"
                     /gene="DNAJA1"
     promoter        3534..3745
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3758..4026
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4061..4753
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="red fluorescent protein from Entacmaea
                     quadricolor"
                     /transl_table=1
                     /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMKIKV
                     VEGGPLPFAFDILATSFMYGSKAFINHTQGIPDFFKQSFPEGFTWERITTYEDGGVLTA
                     TQDTSFQNGCIIYNVKINGVNFPSNGPVMQKKTRGWEANTEMLYPADGGLRGHSQMALK
                     LVGGGYLHCSFKTTYRSKKPAKNLKMPGFHFVDHRLERIKEADKETYVEQHEMAVAKYC
                     DLPSKLGHR"
                     /label="TurboRFP"
     CDS             4754..4807
                     /codon_start=1
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding separate
                     polypeptides."
                     /product="2A peptide from&nbsp;Thosea asigna virus capsid
                     protein"
                     /protein_id=""
                     /transl_table=1
                     /translation="EGRGSLLTCGDVEENPGP"
                     /label="T2A"
     CDS             4808..5407
                     /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    5408..5996
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6070..6303
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6375..6496
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6515..6650
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6688..6704)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6712..6728
                     /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(6736..6766)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6781..6802
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7090..7678)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7849..8709)
                     /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(8710..8814)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9288..9304
                     /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 agagctagcc accatggtga aagaaacaac ttactacgat gttttggggg tcaaacccaa
     2341 tgctactcag gaagaattga aaaaggctta taggaaactg gctttgaagt accatcctga
     2401 taagaaccca aatgaaggag agaagtttaa acagatttct caagcttacg aagttctctc
     2461 tgatgcaaag aaaagggaat tatatgacaa aggaggagaa caggcaatta aagagggtgg
     2521 agcaggtggc ggttttggct cccccatgga catctttgat atgttttttg gaggaggagg
     2581 aaggatgcag agagaaagga gaggtaaaaa tgttgtacat cagctctcag taaccctaga
     2641 agacttatat aatggtgcaa caagaaaact ggctctgcaa aagaatgtga tttgtgacaa
     2701 atgtgaaggt agaggaggta agaaaggagc agtagagtgc tgtcccaatt gccgaggtac
     2761 tggaatgcaa ataagaattc atcagatagg acctggaatg gttcagcaaa ttcagtctgt
     2821 gtgcatggag tgccagggcc atggggagcg gatcagtcct aaagatagat gtaaaagctg
     2881 caacggaagg aagatagttc gagagaagaa aattttagaa gttcatattg acaaaggcat
     2941 gaaagatggc cagaagataa cattccatgg tgaaggagac caagaaccag gactggagcc
     3001 aggcgatatt atcattgtgt tagatcagaa ggaccatgct gtttttactc gacgaggaga
     3061 agaccttttc atgtgtatgg acatacagct cgttgaagca ctgtgtggct tccagaagcc
     3121 aatatctact cttgacaacc gaaccatcgt catcacctct catccaggtc agattgtcaa
     3181 gcatggagat atcaagtgtg tactaaatga aggcatgcca atttatcgta gaccatatga
     3241 aaagggtcgc ctaatcatcg aatttaaggt aaactttcct gagaatggct ttctctctcc
     3301 tgataaactg tctttgctgg aaaaactcct acccgagagg aaggaagtgg aagagactga
     3361 tgagatggac caagtagaac tggtggactt tgatccaaat caggaaagac ggcgccacta
     3421 caatggagaa gcatatgagg atgatgaaca tcatcccaga ggtggtgttc agtgtcagac
     3481 ctcttaagga tccgcggccg caaggatctg cgatcgctcc ggtgcccgtc agtgggcaga
     3541 gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaacgggtgc
     3601 ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt
     3661 tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg
     3721 caacgggttt gccgccagaa cacagctgaa gcttcgaggg gctcgcatct ctccttcacg
     3781 cgcccgccgc cctacctgag gccgccatcc acgccggttg agtcgcgttc tgccgcctcc
     3841 cgcctgtggt gcctcctgaa ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag
     3901 accgggcctt tgtccggcgc tcccttggag cctacctaga ctcagccggc tctccacgct
     3961 ttgcctgacc ctgcttgctc aactctacgt ctttgtttcg ttttctgttc tgcgccgtta
     4021 cagatccaag ctgtgaccgg cgcctacgct agacgccacc atgagcgagc tgatcaagga
     4081 gaacatgcac atgaagctgt acatggaggg caccgtgaac aaccaccact tcaagtgcac
     4141 atccgagggc gaaggcaagc cctacgaggg cacccagacc atgaagatca aggtggtcga
     4201 gggcggccct ctccccttcg ccttcgacat cctggctacc agcttcatgt acggcagcaa
     4261 agccttcatc aaccacaccc agggcatccc cgacttcttt aagcagtcct tccctgaggg
     4321 cttcacatgg gagagaatca ccacatacga agacgggggc gtgctgaccg ctacccagga
     4381 caccagcttc cagaacggct gcatcatcta caacgtcaag atcaacgggg tgaacttccc
     4441 atccaacggc cctgtgatgc agaagaaaac acgcggctgg gaggccaaca ccgagatgct
     4501 gtaccccgct gacggcggcc tgagaggcca cagccagatg gccctgaagc tcgtgggcgg
     4561 gggctacctg cactgctcct tcaagaccac atacagatcc aagaaacccg ctaagaacct
     4621 caagatgccc ggcttccact tcgtggacca cagactggaa agaatcaagg aggccgacaa
     4681 agagacctac gtcgagcagc acgagatggc tgtggccaag tactgcgacc tccctagcaa
     4741 actggggcac agagagggca gaggaagtct tctaacatgc ggtgacgtgg aggagaatcc
     4801 cggccctatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag
     4861 ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga
     4921 tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg
     4981 gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac
     5041 gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt
     5101 gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc
     5161 caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg
     5221 tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc
     5281 cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt
     5341 caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg
     5401 tgcctgaaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     5461 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     5521 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     5581 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     5641 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     5701 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     5761 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
     5821 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     5881 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     5941 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg
     6001 tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa
     6061 aggggggact ggaagggcta attcactccc aacgaaaata agatctgctt tttgcttgta
     6121 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
     6181 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
     6241 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
     6301 gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat
     6361 cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     6421 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     6481 tcatcaatgt atcttatcat gtctggctct agctatcccg cccctaactc cgcccagttc
     6541 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     6601 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc tagacttttg
     6661 cagagacggc ccaaattcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     6721 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     6781 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     6841 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     6901 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     6961 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     7021 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     7081 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     7141 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     7201 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     7261 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     7321 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     7381 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     7441 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     7501 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     7561 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     7621 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     7681 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     7741 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     7801 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     7861 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     7921 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     7981 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     8041 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     8101 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     8161 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     8221 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     8281 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     8341 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     8401 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     8461 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     8521 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     8581 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     8641 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     8701 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     8761 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     8821 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     8881 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     8941 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     9001 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     9061 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
     9121 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga
     9181 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
     9241 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
     9301 cagtgccaag ctg
//
