LOCUS       V043954                 9510 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043954
VERSION     V043954
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..3592
                     /label="GOT2(NM_002080)"
                     /note="GOT2(NM_002080)"
                     /gene="GOT2"
     CDS             3599..3664
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3708..3919
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3932..4200
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4235..4942
                     /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             4952..5005
                     /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             5006..5605
                     /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    5606..6193
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6267..6500
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6572..6693
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6712..6847
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6885..6901)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6909..6925
                     /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(6933..6963)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6978..6999
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7287..7875)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8046..8906)
                     /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(8907..9011)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9485..9501
                     /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 ccatggccct gctgcactcc ggccgcgtcc tccccgggat
     2341 cgccgccgcc ttccacccgg gcctcgccgc cgcggcctct gccagagcca gctcctggtg
     2401 gacccatgtg gaaatgggac ctccagatcc cattctggga gtcactgaag cctttaagag
     2461 ggacaccaat agcaaaaaga tgaatctggg agttggtgcc taccgggatg ataatggaaa
     2521 gccttacgtt ctgcctagcg tccgcaaggc agaggcccag attgccgcaa aaaatttgga
     2581 caaggaatac ctgcccattg ggggactggc tgaattttgc aaggcatctg cagaactagc
     2641 cctgggtgag aacagcgaag tcttgaagag tggccggttt gtcactgtgc agaccatttc
     2701 tggaactgga gccttaagga tcggagccag ttttctgcaa agatttttta agttcagccg
     2761 agatgtcttt ctgcccaaac caacctgggg aaaccacaca cccatcttca gggatgctgg
     2821 catgcagcta caaggttatc ggtattatga ccccaagact tgcggttttg acttcacagg
     2881 cgctgtggag gatatttcaa aaataccaga gcagagtgtt cttcttctgc atgcctgcgc
     2941 ccacaatccc acgggagtgg acccgcgtcc ggaacagtgg aaggaaatag caacagtggt
     3001 gaagaaaagg aatctctttg cgttctttga catggcctac caaggctttg ccagtggtga
     3061 tggtgataag gatgcctggg ctgtgcgcca cttcatcgaa cagggcatta atgtttgcct
     3121 ctgccaatca tatgccaaga acatgggctt atatggtgag cgtgtaggag ccttcactat
     3181 ggtctgcaaa gatgcggatg aagccaaaag ggtagagtca cagttgaaga tcttgatccg
     3241 tcccatgtat tccaaccctc ccctcaatgg ggcccggatt gctgctgcca ttctgaacac
     3301 cccagatttg cgaaaacaat ggctgcaaga agtgaaagtc atggctgacc gcatcattgg
     3361 catgcggact caactggtct ccaacctcaa gaaggagggt tccacccaca attggcaaca
     3421 catcaccgac caaattggca tgttctgttt cacagggcta aagcctgaac aggtggagcg
     3481 gctgatcaag gagttctcca tctacatgac aaaagatggc cgcatctctg tggcaggggt
     3541 cacctccagc aacgtgggct accttgccca tgccattcac caggtcacca agggatccga
     3601 ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg atgacgatga
     3661 caagtagcgg ccgcgaagga tctgcgatcg ctccggtgcc cgtcagtggg cagagcgcac
     3721 atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgaacgg gtgcctagag
     3781 aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga
     3841 gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg
     3901 gtttgccgcc agaacacagc tgaagcttcg aggggctcgc atctctcctt cacgcgcccg
     3961 ccgccctacc tgaggccgcc atccacgccg gttgagtcgc gttctgccgc ctcccgcctg
     4021 tggtgcctcc tgaactgcgt ccgccgtcta ggtaagttta aagctcaggt cgagaccggg
     4081 cctttgtccg gcgctccctt ggagcctacc tagactcagc cggctctcca cgctttgcct
     4141 gaccctgctt gctcaactct acgtctttgt ttcgttttct gttctgcgcc gttacagatc
     4201 caagctgtga ccggcgccta cgctagacgc caccatggtg agcaagggcg aggaggataa
     4261 catggccatc atcaaggagt tcatgcgctt caaggtgcac atggagggct ccgtgaacgg
     4321 ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc
     4381 caagctgaag gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgtcccctca
     4441 gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa
     4501 gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt
     4561 ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct
     4621 gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaagacca tgggctggga
     4681 ggcctcctcc gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagcagag
     4741 gctgaagctg aaggacggcg gccactacga cgctgaggtc aagaccacct acaaggccaa
     4801 gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc aagttggaca tcacctccca
     4861 caacgaggac tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg
     4921 cggcatggac gagctgtaca agggatctcg cgagggcaga ggaagtcttc taacatgcgg
     4981 tgacgtggag gagaatcccg gccctatgac cgagtacaag cccacggtgc gcctcgccac
     5041 ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc
     5101 cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact
     5161 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc
     5221 ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg
     5281 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct
     5341 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc
     5401 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga
     5461 gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga
     5521 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg
     5581 catgacccgc aagcccggtg cctgaaatca acctctggat tacaaaattt gtgaaagatt
     5641 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     5701 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     5761 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     5821 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     5881 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     5941 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     6001 gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     6061 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     6121 ggctctgcgg cctcttccgc gtctccgcct tcgccctcag acgagtcgga tctccctttg
     6181 gccgcctccc cgcctggtac ctttaagacc aatgacttac aaggcagctg tagatcttag
     6241 ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac gaaaataaga
     6301 tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc
     6361 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
     6421 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
     6481 agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt atttataact
     6541 tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt ataatggtta
     6601 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     6661 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc tatcccgccc
     6721 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     6781 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     6841 ggaggcctag acttttgcag agacggccca aattcgtaat catggtcata gctgtttcct
     6901 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     6961 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     7021 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     7081 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     7141 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     7201 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     7261 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     7321 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     7381 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     7441 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     7501 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     7561 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     7621 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     7681 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     7741 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     7801 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     7861 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     7921 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     7981 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     8041 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     8101 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     8161 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     8221 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     8281 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     8341 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     8401 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     8461 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     8521 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     8581 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     8641 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     8701 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     8761 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     8821 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     8881 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     8941 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     9001 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     9061 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt
     9121 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     9181 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     9241 ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt
     9301 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc gccattcagg
     9361 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg
     9421 aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
     9481 cgttgtaaaa cgacggccag tgccaagctg
//
