LOCUS       V027882                 9331 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027882
VERSION     V027882
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..3357
                     /label="LRG1(NM_052972)"
                     /note="LRG1(NM_052972)"
                     /gene="LRG1"
     CDS             3364..3429
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3473..3684
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3697..3965
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      3994..4003
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             4024..4686
                     /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             4756..4809
                     /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             4810..5409
                     /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    5410..5998
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6072..6305
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6377..6498
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6538..6673
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6706..6722)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6730..6746
                     /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(6754..6784)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6799..6820
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7108..7696)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7867..8727)
                     /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(8728..8832)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9306..9322
                     /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 cctcttggag cagacagcga
     2341 ccaaaaagcc cagggggcat tcaaccccat gtttctagaa ctctgttcct gctgctgctg
     2401 ttggcagcct cagcctgggg ggtcaccctg agccccaaag actgccaggt gttccgctca
     2461 gaccatggca gctccatctc ctgtcaacca cctgccgaaa tccccggcta cctgccagcc
     2521 gacaccgtgc acctggccgt ggaattcttc aacctgaccc acctgccagc caacctcctc
     2581 cagggcgcct ctaagctcca agaattgcac ctctccagca atgggctgga aagcctctcg
     2641 cccgaattcc tgcggccagt gccgcagctg agggtgctgg atctaacccg aaacgccctg
     2701 accgggctgc ccccgggcct cttccaggcc tcagccaccc tggacaccct ggtattgaaa
     2761 gaaaaccagc tggaggtcct ggaggtctcg tggctacacg gcctgaaagc tctggggcat
     2821 ctggacctgt ctgggaaccg cctccggaaa ctgccccccg ggctgctggc caacttcacc
     2881 ctcctgcgca cccttgacct tggggagaac cagttggaga ccttgccacc tgacctcctg
     2941 aggggtccgc tgcaattaga acggctacat ctagaaggca acaaattgca agtactggga
     3001 aaagatctcc tcttgccgca gccggacctg cgctacctct tcctgaacgg caacaagctg
     3061 gccagggtgg cagccggtgc cttccagggc ctgcggcagc tggacatgct ggacctctcc
     3121 aataactcac tggccagcgt gcccgagggg ctctgggcat ccctagggca gccaaactgg
     3181 gacatgcggg atggcttcga catctccggc aacccctgga tctgtgacca gaacctgagc
     3241 gacctctatc gttggcttca ggcccaaaaa gacaagatgt tttcccagaa tgacacgcgc
     3301 tgtgctgggc ctgaagccgt gaagggccag acgctcctgg cagtggccaa gtcccaggga
     3361 tccgactaca aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac
     3421 gatgacaagt agcggccgcg aaggatctgc gatcgctccg gtgcccgtca gtgggcagag
     3481 cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg aacgggtgcc
     3541 tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt
     3601 cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc
     3661 aacgggtttg ccgccagaac acagctgaag cttcgagggg ctcgcatctc tccttcacgc
     3721 gcccgccgcc ctacctgagg ccgccatcca cgccggttga gtcgcgttct gccgcctccc
     3781 gcctgtggtg cctcctgaac tgcgtccgcc gtctaggtaa gtttaaagct caggtcgaga
     3841 ccgggccttt gtccggcgct cccttggagc ctacctagac tcagccggct ctccacgctt
     3901 tgcctgaccc tgcttgctca actctacgtc tttgtttcgt tttctgttct gcgccgttac
     3961 agatccaagc tgtgaccggc gcctacgcta gacgccacca tggagagcga cgagagcggc
     4021 ctgcccgcca tggagatcga gtgccgcatc accggcaccc tgaacggcgt ggagttcgag
     4081 ctggtgggcg gcggagaggg cacccccaag cagggccgca tgaccaacaa gatgaagagc
     4141 accaaaggcg ccctgacctt cagcccctac ctgctgagcc acgtgatggg ctacggcttc
     4201 taccacttcg gcacctaccc cagcggctac gagaacccct tcctgcacgc catcaacaac
     4261 ggcggctaca ccaacacccg catcgagaag tacgaggacg gcggcgtgct gcacgtgagc
     4321 ttcagctacc gctacgaggc cggccgcgtg atcggcgact tcaaggtggt gggcaccggc
     4381 ttccccgagg acagcgtgat cttcaccgac aagatcatcc gcagcaacgc caccgtggag
     4441 cacctgcacc ccatgggcga taacgtgctg gtgggcagct tcgcccgcac cttcagcctg
     4501 cgcgacggcg gctactacag cttcgtggtg gacagccaca tgcacttcaa gagcgccatc
     4561 caccccagca tcctgcagaa cgggggcccc atgttcgcct tccgccgcgt ggaggagctg
     4621 cacagcaaca ccgagctggg catcgtggag taccagcacg ccttcaagac ccccatcgcc
     4681 ttcgccagat cccgcgctca gtcgtccaat tctgccgtgg acggcaccgc cggacccggc
     4741 tccaccggat ctcgcgaggg cagaggaagt cttctaacat gcggtgacgt ggaggagaat
     4801 cccggcccta tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc
     4861 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc
     4921 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc
     4981 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc
     5041 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag
     5101 ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg
     5161 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag
     5221 ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc
     5281 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc
     5341 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc
     5401 ggtgcctgaa atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     5461 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     5521 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     5581 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     5641 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     5701 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     5761 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct
     5821 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     5881 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     5941 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct
     6001 ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga
     6061 aaagggggga ctggaagggc taattcactc ccaacgaaga taagatctgc tttttgcttg
     6121 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
     6181 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
     6241 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
     6301 tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat
     6361 atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     6421 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     6481 actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccatc
     6541 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     6601 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     6661 ttttttggag gcctagactt ttgcagagac caaattcgta atcatgtcat agctgtttcc
     6721 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     6781 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     6841 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     6901 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     6961 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     7021 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     7081 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     7141 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     7201 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     7261 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     7321 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     7381 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     7441 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     7501 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     7561 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     7621 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     7681 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     7741 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     7801 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     7861 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     7921 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     7981 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     8041 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     8101 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     8161 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     8221 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     8281 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     8341 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     8401 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     8461 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     8521 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     8581 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     8641 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     8701 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     8761 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     8821 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     8881 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     8941 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     9001 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     9061 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
     9121 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag
     9181 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     9241 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     9301 acgttgtaaa acgacggcca gtgccaagct g
//
