LOCUS       V024765                 9436 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024765
VERSION     V024765
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..3478
                     /label="Pa2g4(NM_011119)"
                     /note="Pa2g4(NM_011119)"
                     /gene="Pa2g4"
     CDS             3485..3550
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3595..3806
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3819..4087
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      4116..4125
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             4146..4808
                     /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             4878..4931
                     /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             4932..5531
                     /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    5532..6119
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6193..6426
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6498..6619
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6638..6773
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6811..6827)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6835..6851
                     /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(6859..6889)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6904..6925
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7213..7801)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7972..8832)
                     /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(8833..8937)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9411..9427
                     /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 gtctatataa gcagagctcg tttagtgaac
     2221 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag attctagagc
     2281 tagcgaattc gccaccatgt cgggcgaaga cgagcagcag gagcaaacta tcgccgagga
     2341 cctggtcgtg accaagtata agatgggggg cgacatcgcc aaccgggtgc ttcgatcttt
     2401 ggtggaggct tccagctcag gtgtgtctgt gctgagcttg tgtgagaaag gtgatgccat
     2461 gattatggaa gagacaggga agatcttcaa gaaggaaaag gagatgaaga aaggtattgc
     2521 ctttcctacc agcatttccg taaataactg tgtgtgtcac ttctcccctt tgaagagtga
     2581 ccaggactat atactcaagg aaggtgactt ggtaaaaatt gaccttgggg ttcatgtgga
     2641 tggcttcatt gccaatgtgg ctcacacttt tgtaattggt gtagctcagg ggacccaggt
     2701 aacagggcgg aaagcagatg tcattaaggc cgctcaccta tgtgctgaag ctgccttacg
     2761 actggtcaaa cctggaaacc agaacacaca agtgactgaa gcatggaaca aggttgctca
     2821 ctcatttaac tgcacaccaa tagaaggtat gctgtcacac caattgaagc agcatgtgat
     2881 tgatggagag aagacgatta tccagaaccc tacagaccag cagaagaagg accatgaaaa
     2941 ggcagaattt gaggtgcatg aggtttatgc tgtagatgtc ctcgtcagct caggagaagg
     3001 caaggccaaa gatgcaggac agagaaccac catctacaag cgagacccct ctaaacaata
     3061 tggcctgaaa atgaaaactt cacgtgcctt tttcagtgag gtggaacggc gttttgatgc
     3121 catgccgttt actttaagag catttgaaga tgagaagaag gctcgaatgg gtgtggtaga
     3181 gtgtgccaaa catgagttac tacagccatt taacgttctc tatgagaagg agggtgaatt
     3241 tgttgcccag tttaaattta cagttctact catgcccaac ggccccatgc ggataaccag
     3301 tggtcccttt gagcctgacc tgtacaagtc tgagatggag gttcaagatg cagagctgaa
     3361 ggctcttctc cagagttctg caagtagaaa aacccagaaa aagaagaaaa agaaggcctc
     3421 caagactgta gagaatgcca ccagtggaga aaccttagaa gagaatggag ctggggacgg
     3481 atccgactac aaagaccatg acggtgatta taaagatcat gacatcgact acaaggatga
     3541 cgatgacaag tgagcggccg cgaaggatct gcgatcgctc cggtgcccgt cagtgggcag
     3601 agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaacgggtg
     3661 cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt
     3721 ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc
     3781 gcaacgggtt tgccgccaga acacagctga agcttcgagg ggctcgcatc tctccttcac
     3841 gcgcccgccg ccctacctga ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc
     3901 ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga
     3961 gaccgggcct ttgtccggcg ctcccttgga gcctacctag actcagccgg ctctccacgc
     4021 tttgcctgac cctgcttgct caactctacg tctttgtttc gttttctgtt ctgcgccgtt
     4081 acagatccaa gctgtgaccg gcgcctacgc tagacgccac catggagagc gacgagagcg
     4141 gcctgcccgc catggagatc gagtgccgca tcaccggcac cctgaacggc gtggagttcg
     4201 agctggtggg cggcggagag ggcaccccca agcagggccg catgaccaac aagatgaaga
     4261 gcaccaaagg cgccctgacc ttcagcccct acctgctgag ccacgtgatg ggctacggct
     4321 tctaccactt cggcacctac cccagcggct acgagaaccc cttcctgcac gccatcaaca
     4381 acggcggcta caccaacacc cgcatcgaga agtacgagga cggcggcgtg ctgcacgtga
     4441 gcttcagcta ccgctacgag gccggccgcg tgatcggcga cttcaaggtg gtgggcaccg
     4501 gcttccccga ggacagcgtg atcttcaccg acaagatcat ccgcagcaac gccaccgtgg
     4561 agcacctgca ccccatgggc gataacgtgc tggtgggcag cttcgcccgc accttcagcc
     4621 tgcgcgacgg cggctactac agcttcgtgg tggacagcca catgcacttc aagagcgcca
     4681 tccaccccag catcctgcag aacgggggcc ccatgttcgc cttccgccgc gtggaggagc
     4741 tgcacagcaa caccgagctg ggcatcgtgg agtaccagca cgccttcaag acccccatcg
     4801 ccttcgccag atcccgcgct cagtcgtcca attctgccgt ggacggcacc gccggacccg
     4861 gctccaccgg atctcgcgag ggcagaggaa gtcttctaac atgcggtgac gtggaggaga
     4921 atcccggccc tatgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc
     4981 ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg
     5041 tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg
     5101 tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga
     5161 ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg
     5221 agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc
     5281 ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca
     5341 agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc
     5401 ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca
     5461 ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc
     5521 ccggtgcctg aaatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     5581 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     5641 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     5701 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     5761 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     5821 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     5881 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc
     5941 catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     6001 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     6061 ttccgcgtct ccgccttcgc cctcagacga gtcggatctc cctttggccg cctccccgcc
     6121 tggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag
     6181 aaaagggggg actggaaggg ctaattcact cccaacgaaa ataagatctg ctttttgctt
     6241 gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga
     6301 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc
     6361 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct
     6421 ctagcagtag tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa
     6481 tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     6541 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     6601 aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa ctccgcccag
     6661 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     6721 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctagactt
     6781 ttgcagagac ggcccaaatt cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     6841 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     6901 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     6961 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     7021 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     7081 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     7141 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     7201 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     7261 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     7321 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     7381 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     7441 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     7501 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     7561 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     7621 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     7681 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     7741 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     7801 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     7861 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     7921 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     7981 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     8041 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     8101 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     8161 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     8221 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     8281 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     8341 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     8401 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     8461 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     8521 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     8581 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     8641 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     8701 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     8761 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     8821 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     8881 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     8941 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     9001 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     9061 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     9121 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     9181 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
     9241 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg
     9301 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     9361 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
     9421 ggccagtgcc aagctg
//
