LOCUS       V025793                 9735 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025793
VERSION     V025793
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     LTR             1..634
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     misc_feature    681..806
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1303..1536
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1721..1765
                     /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    2028..2143
                     /note="central polypurine tract and central termination
                     sequence of HIV-1 (lacking the first T)"
                     /label="cPPT/CTS"
     enhancer        2201..2504
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        2505..2708
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2815..4365
                     /label="USP30(NM_032663)"
                     /note="USP30(NM_032663)"
                     /gene="USP30"
     CDS             4372..4401
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     misc_feature    4438..5024
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
                     /label="IRES2"
     CDS             5044..5643
                     /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    5657..6245
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6452..7085
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7213..7229)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7237..7253
                     /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(7261..7291)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7306..7327
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7615..8203)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8374..9234)
                     /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(9235..9339)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9387..9521
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag
     2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2161 acagcagaga tccagtttat cgataagctt gggagttccg cgttacataa cttacggtaa
     2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc
     2761 tccatagaag acaccgactc tactagagga tctatttccg gtgaattcgc caccatgctg
     2821 agctcccggg ccgaggcggc gatgaccgcg gccgacaggg ccatccagcg cttcctgcgg
     2881 accggggcgg ccgtcagata taaagtcatg aagaactggg gagttatagg tggaattgct
     2941 gctgctcttg cagcaggaat atatgttatt tggggtccca ttacagaaag aaagaagcgt
     3001 agaaaagggc ttgtgcctgg ccttgttaat ttagggaaca cctgcttcat gaactccctg
     3061 ctacaaggcc tgtctgcctg tcctgctttc atcaggtggc tggaagagtt cacctcccag
     3121 tactccaggg atcagaagga gcccccctca caccagtatt tatccttaac actcttgcac
     3181 cttctgaaag ccttgtcctg ccaagaagtt actgatgatg aggtcttaga tgcaagctgc
     3241 ttgttggatg tcttaagaat gtacagatgg cagatctcat catttgaaga acaggatgct
     3301 cacgaattat tccatgtcat tacctcgtca ttggaagatg agcgagaccg ccagcctcgg
     3361 gtcacacatt tgtttgatgt gcattccctg gagcagcagt cagaaataac tcccaaacaa
     3421 attacctgcc gcacaagagg gtcacctcac cctacatcca atcactggaa gtctcaacat
     3481 ccttttcatg gaagactcac tagtaatatg gtctgcaaac actgtgaaca ccagagtcct
     3541 gttcgatttg atacctttga tagcctttca ctaagtattc cagccgccac atggggtcac
     3601 ccattgaccc tggaccactg ccttcaccac ttcatctcat cagaatcagt gcgggatgtt
     3661 gtgtgtgaca actgtacaaa gattgaagcc aagggaacgt tgaacgggga aaaggtggaa
     3721 caccagagga ccacttttgt taaacagtta aaactaggga agctccctca gtgtctctgc
     3781 atccacctac agcggctgag ctggtccagc cacggcacgc ctctgaagcg gcatgagcac
     3841 gtgcagttca atgagttcct gatgatggac atttacaagt accacctcct tggacataaa
     3901 cctagtcaac acaaccctaa actgaacaag aacccagggc ctacactgga gctgcaggat
     3961 gggccgggag cccccacacc agttctgaat cagccagggg cccccaaaac acagattttt
     4021 atgaatggcg cctgctcccc atctttattg ccaacgctgt cagcgccgat gcccttccct
     4081 ctcccagttg ttcccgacta cagctcctcc acatacctct tccggctgat ggcagttgtc
     4141 gtccaccatg gagacatgca ctctggacac tttgtcactt accgacggtc cccaccttct
     4201 gccaggaacc ctctctcaac tagcaatcag tggctgtggg tctccgatga cactgtccgc
     4261 aaggccagcc tgcaggaggt cctgtcctcc agcgcctacc tgctgttcta cgagcgcgtc
     4321 ctttccagga tgcagcacca gagccaggag tgcaagtctg aagaactcga ggaacaaaaa
     4381 ctcatctcag aagaggatct gtgatcgaga ctagttctag agcggccgcg gatcccgccc
     4441 ctctccctcc ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc
     4501 gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa
     4561 acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat
     4621 gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac
     4681 aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg
     4741 cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt
     4801 tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg
     4861 gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc ctcggtgcac
     4921 atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga
     4981 cgtggttttc ctttgaaaaa cacgatgata agcttgccac aacccacaag gagacgacct
     5041 tccatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt cccccgggcc
     5101 gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgacccg
     5161 gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc
     5221 gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg
     5281 gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc
     5341 ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag
     5401 gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg
     5461 ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc
     5521 ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc
     5581 gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc
     5641 tagacgcgtc tggaacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     5701 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     5761 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     5821 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     5881 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     5941 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     6001 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc
     6061 ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     6121 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     6181 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     6241 cccgcctgga attaattctg cagtcgagac ctagaaaaac atggagcaat cacaagtagc
     6301 aatacagcag ctaccaatgc tgattgtgcc tggctagaag cacaagagga ggaggaggtg
     6361 ggttttccag tcacacctca ggtaccttta agaccaatga cttacaaggc agctgtagat
     6421 cttagccact ttttaaaaga aaagagggga ctggaagggc taattcactc ccaacgaaga
     6481 caagatatcc ttgatctgtg gatctaccac acacaaggct acttccctga ttagcagaac
     6541 tacacaccag ggccaggggt cagatatcca ctgacctttg gatggtgcta caagctagta
     6601 ccagttgagc cagataaggt agaagaggcc aataaaggag agaacaccag cttgttacac
     6661 cctgtgagcc tgcatgggat ggatgacccg gagagagaag tgttagagtg gaggtttgac
     6721 agccgcctag catttcatca cgtggcccga gagctgcatc cggagtactt caagaactgc
     6781 tgatatcgag cttgctacaa gggactttcc gctggggact ttccagggag gcgtggcctg
     6841 ggcgggactg gggagtggcg agccctcaga tcctgcatat aagcagctgc tttttgcctg
     6901 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
     6961 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
     7021 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
     7081 tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat
     7141 atcagagagt gagaggcctt gacattgcta gcgtttaccg tcgacctcta gctagagctt
     7201 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     7261 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     7321 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     7381 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     7441 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     7501 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     7561 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     7621 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     7681 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     7741 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     7801 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     7861 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     7921 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     7981 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     8041 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     8101 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     8161 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     8221 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     8281 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     8341 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     8401 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     8461 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     8521 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     8581 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     8641 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     8701 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     8761 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     8821 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     8881 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     8941 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     9001 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     9061 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     9121 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     9181 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     9241 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     9301 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     9361 acctgacgtc gacggatcgg gagatcaact tgtttattgc agcttataat ggttacaaat
     9421 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     9481 gtttgtccaa actcatcaat gtatcttatc atgtctggat caactggata actcaagcta
     9541 accaaaatca tcccaaactt cccaccccat accctattac cactgccaat tacctgtggt
     9601 ttcatttact ctaaacctgt gattcctctg aattattttc attttaaaga aattgtattt
     9661 gttaaatatg tactacaaac ttagtagttt ttaaagaaat tgtatttgtt aaatatgtac
     9721 tacaaactta gtagt
//
