LOCUS       V040928                 9553 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V040928
VERSION     V040928
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             2303..3661
                     /label="GPR39(NM_001508)"
                     /note="GPR39(NM_001508)"
                     /gene="GPR39"
     promoter        3712..3923
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3936..4204
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      4233..4242
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             4263..4925
                     /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             4995..5048
                     /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             5049..5648
                     /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    5649..6236
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6310..6543
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6615..6736
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6755..6890
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6928..6944)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6952..6968
                     /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(6976..7006)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7021..7042
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7330..7918)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8089..8949)
                     /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(8950..9054)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9528..9544
                     /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 gaattcgcca ccatggcttc acccagcctc ccgggcagtg actgctccca
     2341 aatcattgat cacagtcatg tccccgagtt tgaggtggcc acctggatca aaatcaccct
     2401 tattctggtg tacctgatca tcttcgtgat gggccttctg gggaacagcg ccaccattcg
     2461 ggtcacccag gtgctgcaga agaaaggata cttgcagaag gaggtgacag accacatggt
     2521 gagtttggct tgctcggaca tcttggtgtt cctcatcggc atgcccatgg agttctacag
     2581 catcatctgg aatcccctga ccacgtccag ctacaccctg tcctgcaagc tgcacacttt
     2641 cctcttcgag gcctgcagct acgctacgct gctgcacgtg ctgacactca gctttgagcg
     2701 ctacatcgcc atctgtcacc ccttcaggta caaggctgtg tcgggacctt gccaggtgaa
     2761 gctgctgatt ggcttcgtct gggtcacctc cgccctggtg gcactgccct tgctgtttgc
     2821 catgggtact gagtaccccc tggtgaacgt gcccagccac cggggtctca cttgcaaccg
     2881 ctccagcacc cgccaccacg agcagcccga gacctccaat atgtccatct gtaccaacct
     2941 ctccagccgc tggaccgtgt tccagtccag catcttcggc gccttcgtgg tctacctcgt
     3001 ggtcctgctc tccgtagcct tcatgtgctg gaacatgatg caggtgctca tgaaaagcca
     3061 gaagggctcg ctggccgggg gcacgcggcc tccgcagctg aggaagtccg agagcgaaga
     3121 gagcaggacc gccaggaggc agaccatcat cttcctgagg ctgattgttg tgacattggc
     3181 cgtatgctgg atgcccaacc agattcggag gatcatggct gcggccaaac ccaagcacga
     3241 ctggacgagg tcctacttcc gggcgtacat gatcctcctc cccttctcgg agacgttttt
     3301 ctacctcagc tcggtcatca acccgctcct gtacacggtg tcctcgcagc agtttcggcg
     3361 ggtgttcgtg caggtgctgt gctgccgcct gtcgctgcag cacgccaacc acgagaagcg
     3421 cctgcgcgta catgcgcact ccaccaccga cagcgcccgc tttgtgcagc gcccgttgct
     3481 cttcgcgtcc cggcgccagt cctctgcaag gagaactgag aagattttct taagcacttt
     3541 tcagagcgag gccgagcccc agtctaagtc ccagtcattg agtctcgagt cactagagcc
     3601 caactcaggc gcgaaaccag ccaattctgc tgcagagaat ggttttcagg agcatgaagt
     3661 ttgaggatcc gcggccgcga aggatctgcg atcgctccgg tgcccgtcag tgggcagagc
     3721 gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga acgggtgcct
     3781 agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc
     3841 ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca
     3901 acgggtttgc cgccagaaca cagctgaagc ttcgaggggc tcgcatctct ccttcacgcg
     3961 cccgccgccc tacctgaggc cgccatccac gccggttgag tcgcgttctg ccgcctcccg
     4021 cctgtggtgc ctcctgaact gcgtccgccg tctaggtaag tttaaagctc aggtcgagac
     4081 cgggcctttg tccggcgctc ccttggagcc tacctagact cagccggctc tccacgcttt
     4141 gcctgaccct gcttgctcaa ctctacgtct ttgtttcgtt ttctgttctg cgccgttaca
     4201 gatccaagct gtgaccggcg cctacgctag acgccaccat ggagagcgac gagagcggcc
     4261 tgcccgccat ggagatcgag tgccgcatca ccggcaccct gaacggcgtg gagttcgagc
     4321 tggtgggcgg cggagagggc acccccaagc agggccgcat gaccaacaag atgaagagca
     4381 ccaaaggcgc cctgaccttc agcccctacc tgctgagcca cgtgatgggc tacggcttct
     4441 accacttcgg cacctacccc agcggctacg agaacccctt cctgcacgcc atcaacaacg
     4501 gcggctacac caacacccgc atcgagaagt acgaggacgg cggcgtgctg cacgtgagct
     4561 tcagctaccg ctacgaggcc ggccgcgtga tcggcgactt caaggtggtg ggcaccggct
     4621 tccccgagga cagcgtgatc ttcaccgaca agatcatccg cagcaacgcc accgtggagc
     4681 acctgcaccc catgggcgat aacgtgctgg tgggcagctt cgcccgcacc ttcagcctgc
     4741 gcgacggcgg ctactacagc ttcgtggtgg acagccacat gcacttcaag agcgccatcc
     4801 accccagcat cctgcagaac gggggcccca tgttcgcctt ccgccgcgtg gaggagctgc
     4861 acagcaacac cgagctgggc atcgtggagt accagcacgc cttcaagacc cccatcgcct
     4921 tcgccagatc ccgcgctcag tcgtccaatt ctgccgtgga cggcaccgcc ggacccggct
     4981 ccaccggatc tcgcgagggc agaggaagtc ttctaacatg cggtgacgtg gaggagaatc
     5041 ccggccctat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca
     5101 gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg
     5161 atccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg
     5221 ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca
     5281 cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt
     5341 tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc
     5401 ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg
     5461 gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg
     5521 ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg
     5581 tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg
     5641 gtgcctgaaa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     5701 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     5761 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     5821 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     5881 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
     5941 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     6001 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat
     6061 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     6121 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
     6181 cgcgtctccg ccttcgccct cagacgagtc ggatctccct ttggccgcct ccccgcctgg
     6241 tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa
     6301 aggggggact ggaagggcta attcactccc aacgaaaata agatctgctt tttgcttgta
     6361 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
     6421 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
     6481 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
     6541 gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat
     6601 cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     6661 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     6721 tcatcaatgt atcttatcat gtctggctct agctatcccg cccctaactc cgcccagttc
     6781 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     6841 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc tagacttttg
     6901 cagagacggc ccaaattcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     6961 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     7021 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     7081 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     7141 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     7201 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     7261 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     7321 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     7381 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     7441 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     7501 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     7561 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     7621 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     7681 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     7741 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     7801 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     7861 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     7921 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     7981 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     8041 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     8101 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     8161 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     8221 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     8281 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     8341 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     8401 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     8461 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     8521 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     8581 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     8641 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     8701 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     8761 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     8821 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     8881 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     8941 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     9001 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     9061 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     9121 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     9181 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     9241 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     9301 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
     9361 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga
     9421 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
     9481 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
     9541 cagtgccaag ctg
//
