LOCUS       V047612                 9695 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V047612
VERSION     V047612
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..1920
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2010..2213
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2302..3876
                     /label="USP22(NM_015276)"
                     /note="USP22(NM_015276)"
                     /gene="USP22"
     CDS             3913..4623
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /transl_table=1
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK*"
                     /label="mCherry"
     promoter        4665..4876
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4889..5157
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5184..5783
                     /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    5790..6378
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6452..6685
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6757..6878
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6897..7032
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7070..7086)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7094..7110
                     /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(7118..7148)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7163..7184
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7472..8060)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8231..9091)
                     /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(9092..9196)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9670..9686
                     /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 attacaaaat tcaaaatttt
     1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag
     2221 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct
     2281 agagctagcg aattcgccac catggtgtcc cggccagagc ccgagggcga ggccatggac
     2341 gccgagctgg cggtagcgcc gccgggctgc tcgcacctgg gcagcttcaa ggtggacaac
     2401 tggaagcaga acctgcgggc catctaccag tgcttcgtgt ggagcggcac ggctgaggcc
     2461 cgcaagcgca aggccaagtc ctgtatctgc catgtctgtg gcgtccacct caacaggctg
     2521 cattcctgcc tctactgtgt cttcttcggc tgtttcacaa agaagcatat tcacgagcat
     2581 gcgaaggcga agcggcacaa cctggccatt gatctgatgt acggaggcat ctactgtttt
     2641 ctgtgccagg actacatcta tgacaaagac atggaaataa tcgccaagga ggagcagcga
     2701 aaagcttgga aaatgcaagg cgttggagag aagttttcaa cttgggaacc aaccaaacgg
     2761 gagcttgaac tgctgaagca caacccgaaa aggagaaaga tcacctcgaa ctgcaccata
     2821 ggtctgcgtg ggctgatcaa ccttgggaac acatgcttca tgaactgcat cgtgcaggcc
     2881 ctgacccaca cgccacttct gcgggacttc ttcctgtctg acaggcaccg ctgtgagatg
     2941 cagagcccca gctcctgtct ggtctgtgag atgtcctcac tgtttcagga gttttactct
     3001 ggacaccggt cccctcacat cccgtataag ttgctgcacc tggtgtggac ccacgcgagg
     3061 cacctagcag gctacgagca gcaggacgcc cacgagttcc tcatcgcggc cctggacgtg
     3121 ctccaccgac actgcaaagg tgatgacaat gggaagaagg ccaacaaccc caaccactgc
     3181 aactgcatca tagaccagat cttcacaggc gggttgcagt cagacgtcac ctgccaagtc
     3241 tgccatggag tctccaccac catcgacccc ttctgggaca tcagcttgga tctccccggc
     3301 tcttccaccc cattctggcc cctgagccca gggagcgagg gcaacgtggt aaacggggaa
     3361 agccacgtgt cgggaaccac cacgctcacg gactgcctgc gacgattcac cagaccagag
     3421 cacttgggca gcagcgccaa gatcaagtgc agcggttgcc atagctacca ggagtccaca
     3481 aagcagctca ctatgaagaa actgcccatc gtagcctgtt ttcatctcaa acgatttgaa
     3541 cactcagcca agctgcggcg gaagatcacc acgtatgtgt ccttccccct ggagctggac
     3601 atgacccctt tcatggcctc cagcaaagag agcaggatga atggacagta ccagcagccc
     3661 acggacagtc tcaacaatga caacaagtat tccctgtttg ctgttgttaa ccatcaaggg
     3721 accttggaga gtggccacta caccagcttt atccggcagc acaaagacca gtggttcaag
     3781 tgtgacgatg ccatcatcac caaggccagc atcaaggacg tcctggacag cgaagggtac
     3841 ttgctgttct atcacaaaca gttcctggaa tacgagggat ccggtggagg cggatcagga
     3901 ggcggtggat ctatggtgag caagggcgag gaggataaca tggccatcat caaggagttc
     3961 atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
     4021 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt
     4081 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc
     4141 tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc
     4201 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc
     4261 tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc
     4321 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac
     4381 cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc
     4441 cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc
     4501 gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg
     4561 gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag
     4621 taggcggccg cgaaggatct gcgatcgctc cggtgcccgt cagtgggcag agcgcacatc
     4681 gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaacgggtg cctagagaag
     4741 gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg
     4801 tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt
     4861 tgccgccaga acacagctga agcttcgagg ggctcgcatc tctccttcac gcgcccgccg
     4921 ccctacctga ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc ccgcctgtgg
     4981 tgcctcctga actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga gaccgggcct
     5041 ttgtccggcg ctcccttgga gcctacctag actcagccgg ctctccacgc tttgcctgac
     5101 cctgcttgct caactctacg tctttgtttc gttttctgtt ctgcgccgtt acagatccaa
     5161 gctgtgaccg gcgcctacgc tagatgaccg agtacaagcc cacggtgcgc ctcgccaccc
     5221 gcgacgacgt cccccgggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca
     5281 cgcgccacac cgtcgacccg gaccgccaca tcgagcgggt caccgagctg caagaactct
     5341 tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg
     5401 tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc
     5461 cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc
     5521 tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg
     5581 accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc
     5641 gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc
     5701 ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca
     5761 tgacccgcaa gcccggtgcc tgagtcgaca atcaacctct ggattacaaa atttgtgaaa
     5821 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     5881 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     5941 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     6001 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     6061 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     6121 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     6181 ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     6241 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     6301 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     6361 tttgggccgc ctccccgcct ggtaccttta agaccaatga cttacaaggc agctgtagat
     6421 cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaaa
     6481 taagatctgc tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag
     6541 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
     6601 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
     6661 tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat tcagtattta
     6721 taacttgcaa agaaatgaat atcagagagt gagaggaact tgtttattgc agcttataat
     6781 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     6841 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct ctagctatcc
     6901 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     6961 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     7021 tttttggagg cctagacttt tgcagagacg gcccaaattc gtaatcatgg tcatagctgt
     7081 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     7141 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     7201 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     7261 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     7321 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     7381 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     7441 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     7501 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     7561 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     7621 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     7681 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     7741 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     7801 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     7861 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     7921 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     7981 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     8041 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     8101 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     8161 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     8221 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     8281 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     8341 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     8401 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     8461 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     8521 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     8581 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     8641 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     8701 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     8761 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     8821 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     8881 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     8941 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     9001 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     9061 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     9121 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     9181 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
     9241 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt
     9301 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     9361 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     9421 gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc
     9481 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat
     9541 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
     9601 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt
     9661 cacgacgttg taaaacgacg gccagtgcca agctg
//
