LOCUS       V044696                 8965 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044696
VERSION     V044696
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             2317..2991
                     /label="GSTM3(NM_000849)"
                     /note="GSTM3(NM_000849)"
                     /gene="GSTM3"
     CDS             2998..3063
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3107..3318
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3331..3599
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      3628..3637
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             3658..4320
                     /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             4390..4443
                     /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             4444..5043
                     /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    5044..5632
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5706..5939
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6011..6132
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6172..6307
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6340..6356)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6364..6380
                     /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(6388..6418)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6433..6454
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6742..7330)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7501..8361)
                     /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(8362..8466)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8940..8956
                     /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 ctgatacgaa ctcggaattc gccaccatgt cgtgcgagtc gtctatggtt
     2341 ctcgggtact gggatattcg tgggctggcg cacgccatcc gcctgctcct ggagttcacg
     2401 gatacctctt atgaggagaa acggtacacg tgcggggaag ctcctgacta tgatcgaagc
     2461 caatggctgg atgtgaaatt caagctagac ctggactttc ctaatctgcc ctacctcctg
     2521 gatgggaaga acaagatcac ccagagcaat gccatcttgc gctacatcgc tcgcaagcac
     2581 aacatgtgtg gtgagactga agaagaaaag attcgagtgg acatcataga gaaccaagta
     2641 atggatttcc gcacacaact gataaggctc tgttacagct ctgaccacga aaaactgaag
     2701 cctcagtact tggaagagct acctggacaa ctgaaacaat tctccatgtt tctggggaaa
     2761 ttctcatggt ttgccgggga aaagctcacc tttgtggatt ttctcaccta tgatatcttg
     2821 gatcagaacc gtatatttga ccccaagtgc ctggatgagt tcccaaacct gaaggctttc
     2881 atgtgccgtt ttgaggcttt ggagaaaatc gctgcctact tacagtctga tcagttctgc
     2941 aagatgccca tcaacaacaa gatggcccag tggggcaaca agcctgtatg cggatccgac
     3001 tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga tgacgatgac
     3061 aagtagcggc cgcgaaggat ctgcgatcgc tccggtgccc gtcagtgggc agagcgcaca
     3121 tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaacggg tgcctagaga
     3181 aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag
     3241 ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg
     3301 tttgccgcca gaacacagct gaagcttcga ggggctcgca tctctccttc acgcgcccgc
     3361 cgccctacct gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc tcccgcctgt
     3421 ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc gagaccgggc
     3481 ctttgtccgg cgctcccttg gagcctacct agactcagcc ggctctccac gctttgcctg
     3541 accctgcttg ctcaactcta cgtctttgtt tcgttttctg ttctgcgccg ttacagatcc
     3601 aagctgtgac cggcgcctac gctagacgcc accatggaga gcgacgagag cggcctgccc
     3661 gccatggaga tcgagtgccg catcaccggc accctgaacg gcgtggagtt cgagctggtg
     3721 ggcggcggag agggcacccc caagcagggc cgcatgacca acaagatgaa gagcaccaaa
     3781 ggcgccctga ccttcagccc ctacctgctg agccacgtga tgggctacgg cttctaccac
     3841 ttcggcacct accccagcgg ctacgagaac cccttcctgc acgccatcaa caacggcggc
     3901 tacaccaaca cccgcatcga gaagtacgag gacggcggcg tgctgcacgt gagcttcagc
     3961 taccgctacg aggccggccg cgtgatcggc gacttcaagg tggtgggcac cggcttcccc
     4021 gaggacagcg tgatcttcac cgacaagatc atccgcagca acgccaccgt ggagcacctg
     4081 caccccatgg gcgataacgt gctggtgggc agcttcgccc gcaccttcag cctgcgcgac
     4141 ggcggctact acagcttcgt ggtggacagc cacatgcact tcaagagcgc catccacccc
     4201 agcatcctgc agaacggggg ccccatgttc gccttccgcc gcgtggagga gctgcacagc
     4261 aacaccgagc tgggcatcgt ggagtaccag cacgccttca agacccccat cgccttcgcc
     4321 agatcccgcg ctcagtcgtc caattctgcc gtggacggca ccgccggacc cggctccacc
     4381 ggatctcgcg agggcagagg aagtcttcta acatgcggtg acgtggagga gaatcccggc
     4441 cctatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc
     4501 gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg
     4561 gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc
     4621 gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg
     4681 gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc
     4741 ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag
     4801 gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg
     4861 ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc
     4921 ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc
     4981 gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc
     5041 tgaaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     5101 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     5161 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     5221 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     5281 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     5341 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     5401 ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg
     5461 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     5521 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     5581 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggtacc
     5641 tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg
     5701 gggactggaa gggctaattc actcccaacg aagataagat ctgctttttg cttgtactgg
     5761 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
     5821 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
     5881 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
     5941 tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga
     6001 gagtgagagg aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     6061 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     6121 caatgtatct tatcatgtct ggctctagct atcccgcccc taactccgcc catcccgccc
     6181 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     6241 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     6301 ggaggcctag acttttgcag agaccaaatt cgtaatcatg tcatagctgt ttcctgtgtg
     6361 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     6421 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     6481 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     6541 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     6601 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     6661 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     6721 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     6781 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     6841 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     6901 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     6961 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     7021 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     7081 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     7141 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     7201 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     7261 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     7321 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     7381 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     7441 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     7501 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     7561 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     7621 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     7681 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     7741 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     7801 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     7861 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     7921 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     7981 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     8041 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     8101 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     8161 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     8221 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     8281 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     8341 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     8401 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     8461 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     8521 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga
     8581 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     8641 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg
     8701 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat
     8761 accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg
     8821 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
     8881 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
     8941 taaaacgacg gccagtgcca agctg
//
