LOCUS       V041094                 9274 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041094
VERSION     V041094
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..1921
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2011..2214
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2303..3538
                     /label="WDR4(NM_018669)"
                     /note="WDR4(NM_018669)"
                     /gene="WDR4"
     CDS             3545..3610
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3667..3878
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3891..4159
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4194..4901
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /transl_table=1
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
                     /label="mCherry"
     CDS             4911..4964
                     /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             4965..5363
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /note="confers resistance to blasticidin"
                     /product="blasticidin S deaminase"
                     /transl_table=1
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG*"
                     /label="BSD"
     misc_feature    5370..5957
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6031..6264
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6336..6457
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6476..6611
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6649..6665)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6673..6689
                     /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(6697..6727)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6742..6763
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7051..7639)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7810..8670)
                     /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(8671..8775)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9249..9265
                     /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 attacaaaaa ttcaaaattt
     1921 tatcgatact agtattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     1981 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
     2041 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
     2101 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     2161 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggttt atataagcag agctcgttta
     2221 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagattc
     2281 tagagctagc gaattcgcca ccatggcggg ctctgtggga ctggcgttgt gcgggcagac
     2341 gttggtggtg cggggcggca gccgattcct ggccacctcc atagcaagca gtgatgatga
     2401 cagcctcttc atctatgact gcagtgctgc agaaaagaag tcacaagaaa ataaagggga
     2461 ggacgcgccc ttggaccagg ggagcggtgc gattctggcg tccaccttct ccaagtctgg
     2521 cagctatttt gctttaaccg atgacagtaa gcgtctgatt cttttccgta caaaaccatg
     2581 gcaatgtctg agtgtcagga ccgtggcaag gaggtgtaca gccctgactt tcatagcctc
     2641 ggaggagaag gtcttggtgg ccgacaagtc tggagacgtc tactcctttt cggtgctgga
     2701 gccacacggg tgtggccgtc tagagctggg gcacctgtct atgctgttag atgtggctgt
     2761 gagtcctgat gaccgcttca tcctcactgc cgaccgggac gagaagatcc gagtcagctg
     2821 ggccgcggcg ccccatagca tcgagtcctt ctgcttgggg cacacagagt ttgtgagccg
     2881 tatctccgtg gtgccaactc agcccgggct gcttctgtcc tcctctgggg acggcaccct
     2941 gaggctctgg gagtacagga gcggccgcca gctgcactgc tgtcacctgg ccagtctgca
     3001 ggagctggtg gacccccagg ccccccagaa gtttgccgcg tccaggattg cattctggtg
     3061 ccaggagaac tgcgtggcgc tcctgtgcga cggcactcct gtggtctaca tcttccagct
     3121 ggacgcccgc agacagcagt tggtgtacag gcagcagctg gcgttccagc accaagtgtg
     3181 ggacgtggct ttcgaggaga cccaggggct gtgggtgctc caggactgcc aggaagcccc
     3241 cctggtgctc tacaggcctg tgggcgacca gtggcagtct gttcctgaga gcaccgtgtt
     3301 aaagaaagtc tctggtgttc ttcgtgggaa ctgggccatg ctggaaggct ctgccggcgc
     3361 agacgccagc ttcagcagtc tctacaaggc cacgttcgac aacgtgacct cctacctgaa
     3421 gaagaaagag gagagactgc agcagcagct agagaagaag cagcggcgcc ggagtccccc
     3481 gcctgggccc gacgggcatg ccaagaagat gagaccgggg gaggcgacgc taagttgcgg
     3541 atccgactac aaagaccatg acggtgatta taaagatcat gacatcgact acaaggatga
     3601 cgatgacaag tagggatcct gtatggcggc cgcgaaggat ctgcgatcgc tccggtgccc
     3661 gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca
     3721 attgaacggg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact
     3781 ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga
     3841 acgttctttt tcgcaacggg tttgccgcca gaacacagct gaagcttcga ggggctcgca
     3901 tctctccttc acgcgcccgc cgccctacct gaggccgcca tccacgccgg ttgagtcgcg
     3961 ttctgccgcc tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa
     4021 agctcaggtc gagaccgggc ctttgtccgg cgctcccttg gagcctacct agactcagcc
     4081 ggctctccac gctttgcctg accctgcttg ctcaactcta cgtctttgtt tcgttttctg
     4141 ttctgcgccg ttacagatcc aagctgtgac cggcgcctac gctagacgcc accatggtga
     4201 gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca
     4261 tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct
     4321 acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct
     4381 gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg
     4441 acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga
     4501 acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt
     4561 tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga
     4621 agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga
     4681 agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca
     4741 agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca
     4801 agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg
     4861 agggccgcca ctccaccggc ggcatggacg agctgtacaa gggatctcgc gagggcagag
     4921 gaagtcttct aacatgcggt gacgtggagg agaatcccgg ccctatggcc aagcctttgt
     4981 ctcaagaaga atccaccctc attgaaagag caacggctac aatcaacagc atccccatct
     5041 ctgaagacta cagcgtcgcc agcgcagctc tctctagcga cggccgcatc ttcactggtg
     5101 tcaatgtata tcattttact gggggacctt gtgcagaact cgtggtgctg ggcactgctg
     5161 ctgctgcggc agctggcaac ctgacttgta tcgtcgcgat cggaaatgag aacaggggca
     5221 tcttgagccc ctgcggacgg tgccgacagg tgcttctcga tctgcatcct gggatcaaag
     5281 ccatagtgaa ggacagtgat ggacagccga cggcagttgg gattcgtgaa ttgctgccct
     5341 ctggttatgt gtgggagggc taagtcgaca atcaacctct ggattacaaa atttgtgaaa
     5401 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     5461 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     5521 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     5581 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     5641 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     5701 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     5761 ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     5821 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     5881 tgccggctct gcggcctctt ccgcgtctcc gccttcgccc tcagacgagt cggatctccc
     5941 tttggccgcc tccccgcctg gtacctttaa gaccaatgac ttacaaggca gctgtagatc
     6001 ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagat
     6061 aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc
     6121 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
     6181 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
     6241 agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat
     6301 aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg
     6361 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     6421 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc
     6481 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     6541 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     6601 ttttggaggc ctagactttt gcagagacgg cccaaattcg taatcatggt catagctgtt
     6661 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     6721 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     6781 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     6841 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     6901 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     6961 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     7021 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     7081 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     7141 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     7201 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     7261 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     7321 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     7381 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     7441 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     7501 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     7561 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     7621 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     7681 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     7741 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     7801 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     7861 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     7921 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     7981 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     8041 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     8101 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     8161 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     8221 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     8281 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     8341 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     8401 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     8461 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     8521 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     8581 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     8641 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     8701 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     8761 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     8821 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     8881 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     8941 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     9001 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     9061 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt
     9121 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     9181 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     9241 acgacgttgt aaaacgacgg ccagtgccaa gctg
//
