LOCUS       V034456                 9343 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034456
VERSION     V034456
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..3996
                     /label="USP21(NM_012475)"
                     /note="USP21(NM_012475)"
                     /gene="USP21"
     CDS             4003..4068
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4112..4323
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4336..4604
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4637..5431
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MAEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     misc_feature    5438..6026
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6100..6333
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6405..6526
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6545..6680
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6718..6734)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6742..6758
                     /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(6766..6796)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6811..6832
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7120..7708)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7879..8739)
                     /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(8740..8844)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9318..9334
                     /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 catgccccag gcctctgagc accgcctggg ccgtacccga
     2341 gagccacctg ttaatatcca gccccgagtg ggatccaagc taccatttgc ccccagggcc
     2401 cgcagcaagg agcgcagaaa cccagcctct gggccaaacc ccatgttacg acctctgcct
     2461 ccccggccag gtctgcctga tgaacggctc aagaaactgg agctgggacg gggacggacc
     2521 tcaggccctc gtcccagagg cccccttcga gcagatcatg gggttcccct gcctggctca
     2581 ccacccccaa cagtggcttt gcctctccca tctcggacca acttagcccg ttccaagtct
     2641 gtgagcagtg gggacttgcg tccaatgggg attgccttgg gagggcaccg tggcaccgga
     2701 gagcttgggg ctgcactgag ccgcttggcc ctccggcctg agccacccac tttgagacgt
     2761 agcacttctc tccgccgcct agggggcttt cctggacccc ctaccctgtt cagcatacgg
     2821 acagagcccc ctgcttccca tggctccttc cacatgatat ccgcccggtc ctctgagcct
     2881 ttctactctg atgacaagat ggctcatcac acactccttc tgggctctgg tcatgttggc
     2941 cttcgaaacc tgggaaacac gtgcttcctg aatgctgtgc tgcagtgtct gagcagcact
     3001 cgacctcttc gggacttctg tctgagaagg gacttccggc aagaggtgcc tggaggaggc
     3061 cgagcccaag agctcactga agcctttgca gatgtgattg gtgccctctg gcaccctgac
     3121 tcctgcgaag ctgtgaatcc tactcgattc cgagctgtct tccagaaata tgttccctcc
     3181 ttctctggat acagccagca ggatgcccaa gagttcctga agctcctcat ggagcggcta
     3241 caccttgaaa tcaaccgccg aggccgccgg gctccaccga tacttgccaa tggtccagtt
     3301 ccctctccac cccgccgagg aggggctctg ctagaagaac ctgagttaag tgatgatgac
     3361 cgagccaacc taatgtggaa acgttacctg gagcgagagg acagcaagat tgtggacctg
     3421 tttgtgggcc agttgaaaag ttgtctcaag tgccaggcct gtgggtatcg ctccacgacc
     3481 ttcgaggttt tttgtgacct gtccctgccc atccccaaga aaggatttgc tgggggcaag
     3541 gtgtctctgc gggattgttt caaccttttc actaaggaag aagagctaga gtcggagaat
     3601 gccccagtgt gtgaccgatg tcggcagaaa actcgaagta ccaaaaagtt gacagtacaa
     3661 agattccctc gaatcctcgt gctccatctg aatcgatttt ctgcctcccg aggctccatc
     3721 aaaaaaagtt cagtaggtgt agactttcca ctgcagcgac tgagcctagg ggactttgcc
     3781 agtgacaaag ccggaagtcc tgtataccag ctgtatgccc tttgcaacca ctcaggcagc
     3841 gtccactatg gccactacac agccctgtgc cggtgccaga ctggttggca tgtctacaat
     3901 gactctcgtg tctcccctgt cagtgaaaac caggtggcat ccagcgaggg ctacgtgctg
     3961 ttctaccaac tgatgcagga gccaccccgg tgcctgggat ccgactacaa agaccatgac
     4021 ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta gcggccgcga
     4081 aggatctgcg atcgctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc
     4141 gagaagttgg ggggaggggt cggcaattga acgggtgcct agagaaggtg gcgcggggta
     4201 aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg
     4261 tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca
     4321 cagctgaagc ttcgaggggc tcgcatctct ccttcacgcg cccgccgccc tacctgaggc
     4381 cgccatccac gccggttgag tcgcgttctg ccgcctcccg cctgtggtgc ctcctgaact
     4441 gcgtccgccg tctaggtaag tttaaagctc aggtcgagac cgggcctttg tccggcgctc
     4501 ccttggagcc tacctagact cagccggctc tccacgcttt gcctgaccct gcttgctcaa
     4561 ctctacgtct ttgtttcgtt ttctgttctg cgccgttaca gatccaagct gtgaccggcg
     4621 cctacgctag gccaccatgg ctgaacaaga tggattgcac gcaggttctc cggccgcttg
     4681 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     4741 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     4801 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     4861 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     4921 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     4981 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     5041 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     5101 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     5161 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     5221 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     5281 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     5341 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     5401 cttctatcgc cttcttgacg agttcttctg agtcgacaat caacctctgg attacaaaat
     5461 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     5521 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     5581 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     5641 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     5701 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     5761 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     5821 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct
     5881 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     5941 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     6001 gatctccctt tgggccgcct ccccgcctgg tacctttaag accaatgact tacaaggcag
     6061 ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc
     6121 aacgaaaata agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag
     6181 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
     6241 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
     6301 gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc
     6361 agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg tttattgcag
     6421 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     6481 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggctct
     6541 agctatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     6601 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     6661 aggaggcttt tttggaggcc tagacttttg cagagacggc ccaaattcgt aatcatggtc
     6721 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     6781 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     6841 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     6901 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     6961 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     7021 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     7081 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     7141 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     7201 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     7261 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     7321 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     7381 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     7441 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     7501 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     7561 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     7621 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     7681 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     7741 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     7801 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     7861 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     7921 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     7981 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     8041 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     8101 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     8161 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     8221 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     8281 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     8341 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     8401 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     8461 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     8521 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     8581 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     8641 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     8701 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     8761 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     8821 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     8881 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     8941 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     9001 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     9061 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     9121 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca
     9181 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     9241 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     9301 ttcccagtca cgacgttgta aaacgacggc cagtgccaag ctg
//
