LOCUS       V043610                 9233 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043610
VERSION     V043610
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    1798..1915
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        1960..2171
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             2184..2452
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             2498..3301
                     /label="SNAI2(NM_003068)"
                     /note="SNAI2(NM_003068)"
                     /gene="SNAI2"
     CDS             3308..3373
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3392..3891
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             3936..4598
                     /codon_start=1
                     /product="green fluorescent protein 2 from Pontellina
                     plumata, also known as ppluGFP2 (Shagin et al., 2004)"
                     /transl_table=1
                     /translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT
                     FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY
                     EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG
                     YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA"
                     /label="CopGFP"
     CDS             4668..4721
                     /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             4722..5321
                     /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    5328..5916
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5990..6223
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6295..6416
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6435..6570
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6608..6624)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6632..6648
                     /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(6656..6686)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6701..6722
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7010..7598)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7769..8629)
                     /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(8630..8734)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9208..9224
                     /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 ggttaacttt
     1801 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc
     1861 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttatcga
     1921 tactagtaag gatctgcgat cgctccggtg cccgtcagtg ggcagagcgc acatcgccca
     1981 cagtccccga gaagttgggg ggaggggtcg gcaattgaac gggtgcctag agaaggtggc
     2041 gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg
     2101 gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg
     2161 ccagaacaca gctgaagctt cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta
     2221 cctgaggccg ccatccacgc cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct
     2281 cctgaactgc gtccgccgtc taggtaagtt taaagctcag gtcgagaccg ggcctttgtc
     2341 cggcgctccc ttggagccta cctagactca gccggctctc cacgctttgc ctgaccctgc
     2401 ttgctcaact ctacgtcttt gtttcgtttt ctgttctgcg ccgttacaga tccaagctgt
     2461 gaccggcgcc tactctagag ctagcgaatt cgccaccatg ccgcgctcct tcctggtcaa
     2521 gaagcatttc aacgcctcca aaaagccaaa ctacagcgaa ctggacacac atacagtgat
     2581 tatttccccg tatctctatg agagttactc catgcctgtc ataccacaac cagagatcct
     2641 cagctcagga gcatacagcc ccatcactgt gtggactacc gctgctccat tccacgccca
     2701 gctacccaat ggcctctctc ctctttccgg atactcctca tctttggggc gagtgagtcc
     2761 ccctcctcca tctgacacct cctccaagga ccacagtggc tcagaaagcc ccattagtga
     2821 tgaagaggaa agactacagt ccaagctttc agacccccat gccattgaag ctgaaaagtt
     2881 tcagtgcaat ttatgcaata agacctattc aactttttct gggctggcca aacataagca
     2941 gctgcactgc gatgcccagt ctagaaaatc tttcagctgt aaatactgtg acaaggaata
     3001 tgtgagcctg ggcgccctga agatgcatat tcggacccac acattacctt gtgtttgcaa
     3061 gatctgcggc aaggcgtttt ccagaccctg gttgcttcaa ggacacatta gaactcacac
     3121 gggggagaag cctttttctt gccctcactg caacagagca tttgcagaca ggtcaaatct
     3181 gagggctcat ctgcagaccc attctgatgt aaagaaatac cagtgcaaaa actgctccaa
     3241 aaccttctcc agaatgtctc tcctgcacaa acatgaggaa tctggctgct gtgtagcaca
     3301 cctcgaggac tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga
     3361 tgacgatgac aagtagggat ccgcggccgc tgggtagggg aggcgctttt cccaaggcag
     3421 tctggagcat gcgctttagc agccccgctg ggcacttggc gctacacaag tggcctctgg
     3481 cctcgcacac attccacatc caccggtagg cgccaaccgg ctccgttctt tggtggcccc
     3541 ttcgcgccac cttctactcc tcccctagtc aggaagttcc cccccgcccc gcagctcgcg
     3601 tcgtgcagga cgtgacaaat ggaagtagca cgtctcacta gtctcgtgca gatggacagc
     3661 accgctgagc aatggaagcg ggtaggcctt tggggcagcg gccaatagca gctttgctcc
     3721 ttcgctttct gggctcagag gctgggaagg ggtgggtccg ggggcgggct caggggcggg
     3781 ctcaggggcg gggcgggcgc ccgaaggtcc tccggaggcc cggcattctg cacgcttcaa
     3841 aagcgcacgt ctgccgcgct gttctcctct tcctcatctc cgggcctttc gacctgcagc
     3901 ccaagcttac catggagagc gacgagagcg gcctgcccgc catggagatc gagtgccgca
     3961 tcaccggcac cctgaacggc gtggagttcg agctggtggg cggcggagag ggcaccccca
     4021 agcagggccg catgaccaac aagatgaaga gcaccaaagg cgccctgacc ttcagcccct
     4081 acctgctgag ccacgtgatg ggctacggct tctaccactt cggcacctac cccagcggct
     4141 acgagaaccc cttcctgcac gccatcaaca acggcggcta caccaacacc cgcatcgaga
     4201 agtacgagga cggcggcgtg ctgcacgtga gcttcagcta ccgctacgag gccggccgcg
     4261 tgatcggcga cttcaaggtg gtgggcaccg gcttccccga ggacagcgtg atcttcaccg
     4321 acaagatcat ccgcagcaac gccaccgtgg agcacctgca ccccatgggc gataacgtgc
     4381 tggtgggcag cttcgcccgc accttcagcc tgcgcgacgg cggctactac agcttcgtgg
     4441 tggacagcca catgcacttc aagagcgcca tccaccccag catcctgcag aacgggggcc
     4501 ccatgttcgc cttccgccgc gtggaggagc tgcacagcaa caccgagctg ggcatcgtgg
     4561 agtaccagca cgccttcaag acccccatcg ccttcgccag atcccgcgct cagtcgtcca
     4621 attctgccgt ggacggcacc gccggacccg gctccaccgg atctcgcgag ggcagaggaa
     4681 gtcttctaac atgcggtgac gtggaggaga atcccggccc tatgaccgag tacaagccca
     4741 cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt
     4801 tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca
     4861 ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg
     4921 cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg
     4981 tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc
     5041 aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca
     5101 ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg
     5161 gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca
     5221 acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag
     5281 gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg agtcgacaat caacctctgg
     5341 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     5401 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     5461 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
     5521 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
     5581 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
     5641 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     5701 attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca
     5761 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
     5821 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
     5881 agacgagtcg gatctccctt tgggccgcct ccccgcctgg tacctttaag accaatgact
     5941 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta
     6001 attcactccc aacgaaaata agatctgctt tttgcttgta ctgggtctct ctggttagac
     6061 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
     6121 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
     6181 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat
     6241 cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg
     6301 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     6361 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     6421 gtctggctct agctatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     6481 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     6541 agaagtagtg aggaggcttt tttggaggcc tagacttttg cagagacggc ccaaattcgt
     6601 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     6661 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     6721 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     6781 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     6841 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     6901 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     6961 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     7021 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     7081 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     7141 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     7201 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     7261 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     7321 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     7381 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     7441 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     7501 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     7561 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     7621 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     7681 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     7741 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     7801 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     7861 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     7921 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     7981 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     8041 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     8101 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     8161 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     8221 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     8281 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     8341 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     8401 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     8461 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     8521 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     8581 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     8641 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     8701 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     8761 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     8821 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     8881 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     8941 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
     9001 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     9061 tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     9121 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
     9181 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgccaag ctg
//
