LOCUS       V022129                 9334 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022129
VERSION     V022129
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..3448
                     /label="GJA1(NM_000165)"
                     /note="GJA1(NM_000165)"
                     /gene="GJA1"
     promoter        3493..3704
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3717..3985
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      4014..4023
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             4044..4706
                     /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             4776..4829
                     /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             4830..5429
                     /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    5430..6017
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6091..6324
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6396..6517
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6536..6671
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6709..6725)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6733..6749
                     /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(6757..6787)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6802..6823
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7111..7699)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7870..8730)
                     /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(8731..8835)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9309..9325
                     /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 ccatgggtga ctggagcgcc ttaggcaaac tccttgacaa
     2341 ggttcaagcc tactcaactg ctggagggaa ggtgtggctg tcagtacttt tcattttccg
     2401 aatcctgctg ctggggacag cggttgagtc agcctgggga gatgagcagt ctgcctttcg
     2461 ttgtaacact cagcaacctg gttgtgaaaa tgtctgctat gacaagtctt tcccaatctc
     2521 tcatgtgcgc ttctgggtcc tgcagatcat atttgtgtct gtacccacac tcttgtacct
     2581 ggctcatgtg ttctatgtga tgcgaaagga agagaaactg aacaagaaag aggaagaact
     2641 caaggttgcc caaactgatg gtgtcaatgt ggacatgcac ttgaagcaga ttgagataaa
     2701 gaagttcaag tacggtattg aagagcatgg taaggtgaaa atgcgagggg ggttgctgcg
     2761 aacctacatc atcagtatcc tcttcaagtc tatctttgag gtggccttct tgctgatcca
     2821 gtggtacatc tatggattca gcttgagtgc tgtttacact tgcaaaagag atccctgccc
     2881 acatcaggtg gactgtttcc tctctcgccc cacggagaaa accatcttca tcatcttcat
     2941 gctggtggtg tccttggtgt ccctggcctt gaatatcatt gaactcttct atgttttctt
     3001 caagggcgtt aaggatcggg ttaagggaaa gagcgaccct taccatgcga ccagtggtgc
     3061 gctgagccct gccaaagact gtgggtctca aaaatatgct tatttcaatg gctgctcctc
     3121 accaaccgct cccctctcgc ctatgtctcc tcctgggtac aagctggtta ctggcgacag
     3181 aaacaattct tcttgccgca attacaacaa gcaagcaagt gagcaaaact gggctaatta
     3241 cagtgcagaa caaaatcgaa tggggcaggc gggaagcacc atctctaact cccatgcaca
     3301 gccttttgat ttccccgatg ataaccagaa ttctaaaaaa ctagctgctg gacatgaatt
     3361 acagccacta gccattgtgg accagcgacc ttcaagcaga gccagcagtc gtgccagcag
     3421 cagacctcgg cctgatgacc tggagatcta ggcggccgcg aaggatctgc gatcgctccg
     3481 gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg gggggagggg
     3541 tcggcaattg aacgggtgcc tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg
     3601 tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt gcagtagtcg
     3661 ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acagctgaag cttcgagggg
     3721 ctcgcatctc tccttcacgc gcccgccgcc ctacctgagg ccgccatcca cgccggttga
     3781 gtcgcgttct gccgcctccc gcctgtggtg cctcctgaac tgcgtccgcc gtctaggtaa
     3841 gtttaaagct caggtcgaga ccgggccttt gtccggcgct cccttggagc ctacctagac
     3901 tcagccggct ctccacgctt tgcctgaccc tgcttgctca actctacgtc tttgtttcgt
     3961 tttctgttct gcgccgttac agatccaagc tgtgaccggc gcctacgcta gacgccacca
     4021 tggagagcga cgagagcggc ctgcccgcca tggagatcga gtgccgcatc accggcaccc
     4081 tgaacggcgt ggagttcgag ctggtgggcg gcggagaggg cacccccaag cagggccgca
     4141 tgaccaacaa gatgaagagc accaaaggcg ccctgacctt cagcccctac ctgctgagcc
     4201 acgtgatggg ctacggcttc taccacttcg gcacctaccc cagcggctac gagaacccct
     4261 tcctgcacgc catcaacaac ggcggctaca ccaacacccg catcgagaag tacgaggacg
     4321 gcggcgtgct gcacgtgagc ttcagctacc gctacgaggc cggccgcgtg atcggcgact
     4381 tcaaggtggt gggcaccggc ttccccgagg acagcgtgat cttcaccgac aagatcatcc
     4441 gcagcaacgc caccgtggag cacctgcacc ccatgggcga taacgtgctg gtgggcagct
     4501 tcgcccgcac cttcagcctg cgcgacggcg gctactacag cttcgtggtg gacagccaca
     4561 tgcacttcaa gagcgccatc caccccagca tcctgcagaa cgggggcccc atgttcgcct
     4621 tccgccgcgt ggaggagctg cacagcaaca ccgagctggg catcgtggag taccagcacg
     4681 ccttcaagac ccccatcgcc ttcgccagat cccgcgctca gtcgtccaat tctgccgtgg
     4741 acggcaccgc cggacccggc tccaccggat ctcgcgaggg cagaggaagt cttctaacat
     4801 gcggtgacgt ggaggagaat cccggcccta tgaccgagta caagcccacg gtgcgcctcg
     4861 ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc
     4921 ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag
     4981 aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg
     5041 ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga
     5101 tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag
     5161 gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct
     5221 cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg
     5281 ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct
     5341 acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct
     5401 ggtgcatgac ccgcaagccc ggtgcctgaa atcaacctct ggattacaaa atttgtgaaa
     5461 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     5521 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     5581 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     5641 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     5701 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     5761 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     5821 ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     5881 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     5941 tgccggctct gcggcctctt ccgcgtctcc gccttcgccc tcagacgagt cggatctccc
     6001 tttggccgcc tccccgcctg gtacctttaa gaccaatgac ttacaaggca gctgtagatc
     6061 ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaaaat
     6121 aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc
     6181 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
     6241 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
     6301 agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat
     6361 aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg
     6421 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     6481 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc
     6541 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     6601 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     6661 ttttggaggc ctagactttt gcagagacgg cccaaattcg taatcatggt catagctgtt
     6721 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     6781 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     6841 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     6901 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     6961 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     7021 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     7081 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     7141 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     7201 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     7261 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     7321 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     7381 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     7441 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     7501 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     7561 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     7621 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     7681 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     7741 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     7801 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     7861 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     7921 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     7981 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     8041 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     8101 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     8161 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     8221 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     8281 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     8341 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     8401 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     8461 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     8521 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     8581 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     8641 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     8701 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     8761 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     8821 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     8881 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     8941 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     9001 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     9061 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     9121 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt
     9181 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     9241 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     9301 acgacgttgt aaaacgacgg ccagtgccaa gctg
//
