LOCUS       V035411                 9181 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035411
VERSION     V035411
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        2005..2208
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2297..3223
                     /label="Ppp2ca(NM_017039)"
                     /note="Ppp2ca(NM_017039)"
                     /gene="Ppp2ca"
     CDS             3230..3295
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3340..3551
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3564..3832
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      3861..3870
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             3891..4553
                     /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             4623..4676
                     /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             4677..5276
                     /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    5277..5864
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5938..6171
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6243..6364
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6383..6518
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6556..6572)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6580..6596
                     /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(6604..6634)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6649..6670
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6958..7546)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7717..8577)
                     /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(8578..8682)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9156..9172
                     /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 tactagtatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     1981 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
     2041 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     2101 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
     2161 caaatgggcg gtaggcgtgt acggtgggag gtttatataa gcagagctcg tttagtgaac
     2221 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag attctagagc
     2281 tagcgaattc gccaccatgg acgagaagtt gttcaccaag gagctggacc agtggatcga
     2341 gcagctgaac gagtgcaagc agctctccga gtcccaggtc aagagcctct gcgagaaggc
     2401 taaagaaatc ctgacaaaag aatctaatgt tcaggaggtt cgatgtccag tcactgtgtg
     2461 tggagatgtg catgggcaat ttcatgacct catggaactc tttagaattg gtggtaaatc
     2521 accagataca aattacttgt ttatgggaga ctatgtggac agaggatatt actcagttga
     2581 aacagttaca ctgcttgtag ctcttaaggt tcgttaccga gagcgtatca ccatactccg
     2641 agggaatcac gagagcagac agatcacaca agtttatggt ttctacgatg agtgtttaag
     2701 gaaatacgga aatgcaaatg tttggaaata cttcacagac ctttttgact accttcctct
     2761 cactgccttg gtggatgggc agatcttctg tctacatggt ggtctttcac catccataga
     2821 cacactggat cacatccgag cacttgatcg cctacaagaa gttcctcatg agggtccaat
     2881 gtgtgacttg ctgtggtcag atccagatga ccgtggtggc tgggggatat ctcctcgggg
     2941 agctggttat acctttggcc aagatatttc tgagacattt aatcatgcca atggcctcac
     3001 gttggtgtcc agagctcacc agctggtgat ggagggatat aactggtgcc atgaccggaa
     3061 tgtagtaaca attttcagtg ctccaaacta ttgctatcgt tgtggtaacc aagctgcaat
     3121 catggaactt gatgacactc ttaagtattc tttcttgcag ttcgatccag cacctcgtag
     3181 aggcgagcca catgtcactc gtcgtacccc agactacttc ctgggatccg actacaaaga
     3241 ccatgacggt gattataaag atcatgacat cgactacaag gatgacgatg acaagtgagc
     3301 ggccgcgaag gatctgcgat cgctccggtg cccgtcagtg ggcagagcgc acatcgccca
     3361 cagtccccga gaagttgggg ggaggggtcg gcaattgaac gggtgcctag agaaggtggc
     3421 gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg
     3481 gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg
     3541 ccagaacaca gctgaagctt cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta
     3601 cctgaggccg ccatccacgc cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct
     3661 cctgaactgc gtccgccgtc taggtaagtt taaagctcag gtcgagaccg ggcctttgtc
     3721 cggcgctccc ttggagccta cctagactca gccggctctc cacgctttgc ctgaccctgc
     3781 ttgctcaact ctacgtcttt gtttcgtttt ctgttctgcg ccgttacaga tccaagctgt
     3841 gaccggcgcc tacgctagac gccaccatgg agagcgacga gagcggcctg cccgccatgg
     3901 agatcgagtg ccgcatcacc ggcaccctga acggcgtgga gttcgagctg gtgggcggcg
     3961 gagagggcac ccccaagcag ggccgcatga ccaacaagat gaagagcacc aaaggcgccc
     4021 tgaccttcag cccctacctg ctgagccacg tgatgggcta cggcttctac cacttcggca
     4081 cctaccccag cggctacgag aaccccttcc tgcacgccat caacaacggc ggctacacca
     4141 acacccgcat cgagaagtac gaggacggcg gcgtgctgca cgtgagcttc agctaccgct
     4201 acgaggccgg ccgcgtgatc ggcgacttca aggtggtggg caccggcttc cccgaggaca
     4261 gcgtgatctt caccgacaag atcatccgca gcaacgccac cgtggagcac ctgcacccca
     4321 tgggcgataa cgtgctggtg ggcagcttcg cccgcacctt cagcctgcgc gacggcggct
     4381 actacagctt cgtggtggac agccacatgc acttcaagag cgccatccac cccagcatcc
     4441 tgcagaacgg gggccccatg ttcgccttcc gccgcgtgga ggagctgcac agcaacaccg
     4501 agctgggcat cgtggagtac cagcacgcct tcaagacccc catcgccttc gccagatccc
     4561 gcgctcagtc gtccaattct gccgtggacg gcaccgccgg acccggctcc accggatctc
     4621 gcgagggcag aggaagtctt ctaacatgcg gtgacgtgga ggagaatccc ggccctatga
     4681 ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca
     4741 ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc
     4801 acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg
     4861 gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg
     4921 tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc
     4981 ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg
     5041 cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg
     5101 ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga
     5161 cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg
     5221 tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgaaatc
     5281 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     5341 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     5401 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     5461 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     5521 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     5581 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     5641 gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct
     5701 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     5761 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtctccgcc
     5821 ttcgccctca gacgagtcgg atctcccttt ggccgcctcc ccgcctggta cctttaagac
     5881 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg
     5941 aagggctaat tcactcccaa cgaaaataag atctgctttt tgcttgtact gggtctctct
     6001 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     6061 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     6121 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt
     6181 catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga
     6241 ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     6301 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     6361 cttatcatgt ctggctctag ctatcccgcc cctaactccg cccagttccg cccattctcc
     6421 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     6481 gctattccag aagtagtgag gaggcttttt tggaggccta gacttttgca gagacggccc
     6541 aaattcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     6601 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     6661 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     6721 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     6781 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     6841 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     6901 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     6961 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     7021 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     7081 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     7141 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     7201 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     7261 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     7321 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     7381 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     7441 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     7501 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     7561 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     7621 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     7681 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     7741 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     7801 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     7861 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     7921 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     7981 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     8041 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     8101 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     8161 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     8221 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     8281 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     8341 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     8401 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     8461 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     8521 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     8581 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     8641 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     8701 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     8761 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     8821 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     8881 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     8941 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     9001 ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     9061 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     9121 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgccaagct
     9181 g
//
