LOCUS       V045331                 9414 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045331
VERSION     V045331
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..3433
                     /label="MAPK3(NM_002746)"
                     /note="MAPK3(NM_002746)"
                     /gene="MAPK3"
     CDS             3440..3529
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     promoter        3573..3784
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3797..4065
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      4094..4103
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             4124..4786
                     /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             4856..4909
                     /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             4910..5509
                     /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    5510..6097
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6171..6404
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6476..6597
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6616..6751
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6789..6805)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6813..6829
                     /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(6837..6867)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6882..6903
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7191..7779)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7950..8810)
                     /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(8811..8915)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9389..9405
                     /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 gccaccatgg cggcggcggc ggctcagggg ggcgggggcg gggagccccg
     2341 tagaaccgag ggggtcggcc cgggggtccc gggggaggtg gagatggtga aggggcagcc
     2401 gttcgacgtg ggcccgcgct acacgcagtt gcagtacatc ggcgagggcg cgtacggcat
     2461 ggtcagctcg gcctatgacc acgtgcgcaa gactcgcgtg gccatcaaga agatcagccc
     2521 cttcgaacat cagacctact gccagcgcac gctccgggag atccagatcc tgctgcgctt
     2581 ccgccatgag aatgtcatcg gcatccgaga cattctgcgg gcgtccaccc tggaagccat
     2641 gagagatgtc tacattgtgc aggacctgat ggagactgac ctgtacaagt tgctgaaaag
     2701 ccagcagctg agcaatgacc atatctgcta cttcctctac cagatcctgc ggggcctcaa
     2761 gtacatccac tccgccaacg tgctccaccg agatctaaag ccctccaacc tgctcatcaa
     2821 caccacctgc gaccttaaga tttgtgattt cggcctggcc cggattgccg atcctgagca
     2881 tgaccacacc ggcttcctga cggagtatgt ggctacgcgc tggtaccggg ccccagagat
     2941 catgctgaac tccaagggct ataccaagtc catcgacatc tggtctgtgg gctgcattct
     3001 ggctgagatg ctctctaacc ggcccatctt ccctggcaag cactacctgg atcagctcaa
     3061 ccacattctg ggcatcctgg gctccccatc ccaggaggac ctgaattgta tcatcaacat
     3121 gaaggcccga aactacctac agtctctgcc ctccaagacc aaggtggctt gggccaagct
     3181 tttccccaag tcagactcca aagcccttga cctgctggac cggatgttaa cctttaaccc
     3241 caataaacgg atcacagtgg aggaagcgct ggctcacccc tacctggagc agtactatga
     3301 cccgacggat gagccagtgg ccgaggagcc cttcaccttc gccatggagc tggatgacct
     3361 acctaaggag cggctgaagg agctcatctt ccaggagaca gcacgcttcc agcccggagt
     3421 gctggaggcc cccggatccg aacaaaaact aatatcagag gaagatttgg aacagaaact
     3481 aatcagtgaa gaagatttag agcaaaagtt aatttccgag gaggacttat agcggccgcg
     3541 aaggatctgc gatcgctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc
     3601 cgagaagttg gggggagggg tcggcaattg aacgggtgcc tagagaaggt ggcgcggggt
     3661 aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc
     3721 gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac
     3781 acagctgaag cttcgagggg ctcgcatctc tccttcacgc gcccgccgcc ctacctgagg
     3841 ccgccatcca cgccggttga gtcgcgttct gccgcctccc gcctgtggtg cctcctgaac
     3901 tgcgtccgcc gtctaggtaa gtttaaagct caggtcgaga ccgggccttt gtccggcgct
     3961 cccttggagc ctacctagac tcagccggct ctccacgctt tgcctgaccc tgcttgctca
     4021 actctacgtc tttgtttcgt tttctgttct gcgccgttac agatccaagc tgtgaccggc
     4081 gcctacgcta gacgccacca tggagagcga cgagagcggc ctgcccgcca tggagatcga
     4141 gtgccgcatc accggcaccc tgaacggcgt ggagttcgag ctggtgggcg gcggagaggg
     4201 cacccccaag cagggccgca tgaccaacaa gatgaagagc accaaaggcg ccctgacctt
     4261 cagcccctac ctgctgagcc acgtgatggg ctacggcttc taccacttcg gcacctaccc
     4321 cagcggctac gagaacccct tcctgcacgc catcaacaac ggcggctaca ccaacacccg
     4381 catcgagaag tacgaggacg gcggcgtgct gcacgtgagc ttcagctacc gctacgaggc
     4441 cggccgcgtg atcggcgact tcaaggtggt gggcaccggc ttccccgagg acagcgtgat
     4501 cttcaccgac aagatcatcc gcagcaacgc caccgtggag cacctgcacc ccatgggcga
     4561 taacgtgctg gtgggcagct tcgcccgcac cttcagcctg cgcgacggcg gctactacag
     4621 cttcgtggtg gacagccaca tgcacttcaa gagcgccatc caccccagca tcctgcagaa
     4681 cgggggcccc atgttcgcct tccgccgcgt ggaggagctg cacagcaaca ccgagctggg
     4741 catcgtggag taccagcacg ccttcaagac ccccatcgcc ttcgccagat cccgcgctca
     4801 gtcgtccaat tctgccgtgg acggcaccgc cggacccggc tccaccggat ctcgcgaggg
     4861 cagaggaagt cttctaacat gcggtgacgt ggaggagaat cccggcccta tgaccgagta
     4921 caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc
     4981 cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga
     5041 gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt
     5101 gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc
     5161 gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc
     5221 cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt
     5281 cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt
     5341 gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc
     5401 gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt
     5461 gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgaa atcaacctct
     5521 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     5581 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     5641 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     5701 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     5761 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     5821 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     5881 caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc
     5941 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     6001 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtctcc gccttcgccc
     6061 tcagacgagt cggatctccc tttggccgcc tccccgcctg gtacctttaa gaccaatgac
     6121 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct
     6181 aattcactcc caacgaaaat aagatctgct ttttgcttgt actgggtctc tctggttaga
     6241 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
     6301 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
     6361 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca
     6421 tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt
     6481 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     6541 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     6601 tgtctggctc tagctatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     6661 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     6721 cagaagtagt gaggaggctt ttttggaggc ctagactttt gcagagacgg cccaaattcg
     6781 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     6841 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     6901 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     6961 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     7021 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     7081 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     7141 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     7201 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     7261 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     7321 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     7381 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     7441 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     7501 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     7561 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     7621 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     7681 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     7741 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     7801 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     7861 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     7921 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     7981 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     8041 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     8101 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     8161 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     8221 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     8281 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     8341 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     8401 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     8461 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     8521 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     8581 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     8641 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     8701 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     8761 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     8821 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     8881 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     8941 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     9001 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     9061 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     9121 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     9181 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     9241 atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     9301 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
     9361 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gctg
//
