LOCUS       V021057                 9787 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021057
VERSION     V021057
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             2826..3464
                     /label="FAM177A1(NM_001079519)"
                     /note="FAM177A1(NM_001079519)"
                     /gene="FAM177A1"
     regulatory      3495..3504
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             3504..3545
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /transl_table=1
                     /translation="GKPIPNPLLGLDST"
                     /label="V5 tag"
     CDS             3552..4508
                     /codon_start=1
                     /product="mutant of the E. coli biotin ligase BioID
                     engineered for improved catalytic efficiency (Branon et
                     al., 2018)"
                     /transl_table=1
                     /translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW
                     GVDVFTVPGKGYSLPEPIPLLNAKQILGQLDGGSVAVLPVVDSTNQYLLDRIGELKSGD
                     ACIAEYQQAGRGSRGRKWFSPFGANLYLSMFWRLKRGPAAIGLGPVIGIVMAEALRKLG
                     ADKVRVKWPNDLYLQDRKLAGILVELAGITGDAAQIVIGAGINVAMRRVEESVVNQGWI
                     TLQEAGINLDRNTLAATLIRELRAALELFEQEGLAPYLPRWEKLDNFINRPVKLIIGDK
                     EIFGISRGIDKQGALLLEQDGVIKPWMGGEISLRSAEK"
                     /label="TurboID"
     CDS             4515..4541
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein
                     (Fischer et al., 1995)"
                     /transl_table=1
                     /translation="LPPLERLTL"
                     /label="NES"
     promoter        4575..5074
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5095..5694
                     /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    5708..6296
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6503..7136
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7265..7281)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7289..7305
                     /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(7313..7343)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7358..7379
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7667..8255)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8426..9286)
                     /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(9287..9391)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9439..9573
                     /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 tcgctagcgc taccggactc agatctcgag
     2821 ccaccatgga ccaggagcca gtgggcggtg tggaacgagg agaagccgtc gcagcctcgg
     2881 gagctgcggc cgccgcggca ttcggggaat ctgcagggca gatgagtaac gaaagaggct
     2941 ttgaaaatgt agaactggga gtcataggaa aaaagaagaa agtcccaagg agagtcatcc
     3001 actttgttag tggtgaaaca atggaagaat atagcacaga tgaagacgaa gttgatggcc
     3061 tggagaagaa agatgttttg cctactgttg atccgacaaa acttacctgg ggtccctact
     3121 tatggtttta catgcttcgg gctgctacat caactctctc agtgtgtgac ttccttggag
     3181 agaagattgc atctgttttg ggtatcagca ccccaaagta ccaatatgcc attgatgaat
     3241 attatcggat gaagaaggag gaagaagaag aagaagaaga aaacaggatg tctgaagaag
     3301 cagaaaaaca atatcaacag aataaattgc agactgattc cattgttcag acagatcaac
     3361 cagagacagt gatatccagc tcatttgtga atgtcaattt tgaaatggag ggagacagtg
     3421 aagtaattat ggaaagcaag caaaatccag tctctgtccc accaggtggt tctggtggtg
     3481 gttctggtga attcgccacc atgggcaagc ccatccccaa ccccctgctg ggcctggaca
     3541 gcaccgctag caaagacaat actgtgcctc tgaagctgat cgctctcctg gctaatggcg
     3601 agttccatag tggcgaacag ctgggagaaa ccctgggcat gtccagggcc gctatcaaca
     3661 agcacattca gactctgcgc gactggggcg tggacgtgtt caccgtgccc ggaaagggct
     3721 actctctgcc cgagcctatc ccgctgctga acgctaaaca gattctggga cagctggacg
     3781 gcgggagcgt ggcagtcctg cctgtggtcg actccaccaa tcagtacctg ctggatcgaa
     3841 tcggcgagct gaagagtggg gatgcttgca ttgcagaata tcagcaggca gggagaggaa
     3901 gcagagggag gaaatggttc tctccttttg gagctaacct gtacctgagt atgttttggc
     3961 gcctgaagcg gggaccagca gcaatcggcc tgggcccggt catcggaatt gtcatggcag
     4021 aagcgctgcg aaagctggga gcagacaagg tgcgagtcaa atggcccaat gacctgtatc
     4081 tgcaggatag aaagctggca ggcatcctgg tggagctggc cggaataaca ggcgatgctg
     4141 cacagatcgt cattggcgcc gggattaacg tggctatgag gcgcgtggag gaaagcgtgg
     4201 tcaatcaggg ctggatcaca ctgcaggaag cagggattaa cctggacagg aatactctgg
     4261 ccgctacgct gatccgagag ctgcgggcag ccctggaact gttcgagcag gaaggcctgg
     4321 ctccatatct gccacggtgg gagaagctgg ataacttcat caatagaccc gtgaagctga
     4381 tcattgggga caaagagatt ttcgggatta gccgggggat tgataaacag ggagccctgc
     4441 tgctggaaca ggacggagtt atcaaaccct ggatgggcgg agaaatcagt ctgcggtctg
     4501 ccgaaaagct gcagctgcct cccctggagc gcctgaccct ggactaggat cccgcgactc
     4561 tagataattc taccgggtag gggaggcgct tttcccaagg cagtctggag catgcgcttt
     4621 agcagccccg ctgggcactt ggcgctacac aagtggcctc tggcctcgca cacattccac
     4681 atccaccggt aggcgccaac cggctccgtt ctttggtggc cccttcgcgc caccttctac
     4741 tcctccccta gtcaggaagt tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca
     4801 aatggaagta gcacgtctca ctagtctcgt gcagatggac agcaccgctg agcaatggaa
     4861 gcgggtaggc ctttggggca gcggccaata gcagctttgc tccttcgctt tctgggctca
     4921 gaggctggga aggggtgggt ccgggggcgg gctcaggggc gggctcaggg gcggggcggg
     4981 cgcccgaagg tcctccggag gcccggcatt ctgcacgctt caaaagcgca cgtctgccgc
     5041 gctgttctcc tcttcctcat ctccgggcct ttcgacctgc agcccaagct taccatgacc
     5101 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc
     5161 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac
     5221 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc
     5281 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc
     5341 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg
     5401 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg
     5461 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc
     5521 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc
     5581 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc
     5641 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgaccgcgt
     5701 ctggaacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     5761 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     5821 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     5881 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     5941 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     6001 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     6061 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg
     6121 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     6181 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     6241 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg
     6301 aattaattct gcagtcgaga cctagaaaaa catggagcaa tcacaagtag caatacagca
     6361 gctaccaatg ctgattgtgc ctggctagaa gcacaagagg aggaggaggt gggttttcca
     6421 gtcacacctc aggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac
     6481 tttttaaaag aaaagagggg actggaaggg ctaattcact cccaacgaag acaagatatc
     6541 cttgatctgt ggatctacca cacacaaggc tacttccctg attagcagaa ctacacacca
     6601 gggccagggg tcagatatcc actgaccttt ggatggtgct acaagctagt accagttgag
     6661 ccagataagg tagaagaggc caataaagga gagaacacca gcttgttaca ccctgtgagc
     6721 ctgcatggga tggatgaccc ggagagagaa gtgttagagt ggaggtttga cagccgccta
     6781 gcatttcatc acgtggcccg agagctgcat ccggagtact tcaagaactg ctgatatcga
     6841 gcttgctaca agggactttc cgctggggac tttccaggga ggcgtggcct gggcgggact
     6901 ggggagtggc gagccctcag atcctgcata taagcagctg ctttttgcct gtactgggtc
     6961 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
     7021 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
     7081 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtag
     7141 tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa tatcagagag
     7201 tgagaggcct tgacattgct agcgttttac cgtcgacctc tagctagagc ttggcgtaat
     7261 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     7321 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     7381 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     7441 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     7501 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     7561 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     7621 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     7681 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     7741 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     7801 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     7861 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     7921 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     7981 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     8041 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     8101 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     8161 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     8221 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     8281 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     8341 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     8401 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     8461 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     8521 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     8581 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     8641 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     8701 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     8761 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     8821 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     8881 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     8941 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     9001 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     9061 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     9121 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     9181 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     9241 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     9301 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     9361 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     9421 tcgacggatc gggagatcaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     9481 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     9541 aaactcatca atgtatctta tcatgtctgg atcaactgga taactcaagc taaccaaaat
     9601 catcccaaac ttcccacccc ataccctatt accactgcca attacctgtg gtttcattta
     9661 ctctaaacct gtgattcctc tgaattattt tcattttaaa gaaattgtat ttgttaaata
     9721 tgtactacaa acttagtagt ttttaaagaa attgtatttg ttaaatatgt actacaaact
     9781 tagtagt
//
