LOCUS       V044641                 9523 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044641
VERSION     V044641
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"
     regulatory      2296..2305
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             3082..4176
                     /label="HLA-A(NM_002116)"
                     /note="HLA-A(NM_002116)"
                     /gene="HLA-A"
     CDS             4183..4248
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4292..4503
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4516..4784
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4817..5611
                     /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    5618..6206
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6280..6513
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6585..6706
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6725..6860
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6898..6914)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6922..6938
                     /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(6946..6976)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6991..7012
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7300..7888)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8059..8919)
                     /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(8920..9024)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9498..9514
                     /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 catggccacg ggcgcgccct gcaggctagc ctcgagggat
     2341 ccgatatcgc ggccgcttct ttccaaacaa gttttgcctc atgatgtgaa aatgagatac
     2401 catatggatg gatgtgttaa cggtcacagt ttcactattg agggagaagg tgctggtaaa
     2461 ccttatgaag gcaaaaagat cttggaactg cgtgtaacaa agggaggacc acttccattc
     2521 gcctttgaca ttttgtccag cgttttcact tatggtaaca ggtgtttctg tgagtatcca
     2581 gaggatatgc ccgactactt taagcaatca ttgcccgagg gtcactcttg ggagagaact
     2641 ttaatgtttg aggatggagg atgcggtact gcttcagccc atatatcgtt ggacaaaaac
     2701 tgtttcgtgc ataagtctac gtttcatggt gtgaattttc ctgctaatgg tccggtcatg
     2761 cagaaaaagg caatgaattg ggagcctagc tctgaactga tcaccgcatg tgatggtatt
     2821 ctcaaaggcg acgttaccat gtttttgttg ctggaaggtg gtcacagatt aaagtgccaa
     2881 ttcacaacat cctacaaggc tcataaagca gtgaagatgc caccaaatca cattatcgag
     2941 catgtcctag tcaaaaagga agttgctgac ggttttcaaa tccaggaaca cgctgttgca
     3001 aaacacttca cagtagatgt taaggaaact cgcggccgcg aattcagatc tggtggaggc
     3061 ggatcaggag gcggtggatc tatggccgtc atggcgcccc gaaccctcct cctgctactc
     3121 tcgggggccc tggccctgac ccagacctgg gcgggctccc actccatgag gtatttcttc
     3181 acatccgtgt cccggcccgg ccgcggggag ccccgcttca tcgccgtggg ctacgtggac
     3241 gacacgcagt tcgtgcggtt cgacagcgac gccgcgagcc agaggatgga gccgcgggcg
     3301 ccgtggatag agcaggaggg gccggagtat tgggaccagg agacacggaa tgtgaaggcc
     3361 cagtcacaga ctgaccgagt ggacctgggg accctgcgcg gctactacaa ccagagcgag
     3421 gccggttctc acaccatcca gataatgtat ggctgcgacg tggggtcgga cgggcgcttc
     3481 ctccgcgggt accggcagga cgcctacgac ggcaaggatt acatcgccct gaacgaggac
     3541 ctgcgctctt ggaccgcggc ggacatggcg gctcagatca ccaagcgcaa gtgggaggcg
     3601 gcccatgagg cggagcagtt gagagcctac ctggatggca cgtgcgtgga gtggctccgc
     3661 agatacctgg agaacgggaa ggagacgctg cagcgcacgg acccccccaa gacacatatg
     3721 acccaccacc ccatctctga ccatgaggcc accctgaggt gctgggccct gggcttctac
     3781 cctgcggaga tcacactgac ctggcagcgg gatggggagg accagaccca ggacacggag
     3841 ctcgtggaga ccaggcctgc aggggatgga accttccaga agtgggcggc tgtggtggtg
     3901 ccttctggag aggagcagag atacacctgc catgtgcagc atgagggtct gcccaagccc
     3961 ctcaccctga gatgggagct gtcttcccag cccaccatcc ccatcgtggg catcattgct
     4021 ggcctggttc tccttggagc tgtgatcact ggagctgtgg tcgctgccgt gatgtggagg
     4081 aggaagagct cagatagaaa aggagggagt tacactcagg ctgcaagcag tgacagtgcc
     4141 cagggctctg atgtgtccct cacagcttgt aaagtgggat ccgactacaa agaccatgac
     4201 ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta gcggccgcga
     4261 aggatctgcg atcgctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc
     4321 gagaagttgg ggggaggggt cggcaattga acgggtgcct agagaaggtg gcgcggggta
     4381 aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg
     4441 tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca
     4501 cagctgaagc ttcgaggggc tcgcatctct ccttcacgcg cccgccgccc tacctgaggc
     4561 cgccatccac gccggttgag tcgcgttctg ccgcctcccg cctgtggtgc ctcctgaact
     4621 gcgtccgccg tctaggtaag tttaaagctc aggtcgagac cgggcctttg tccggcgctc
     4681 ccttggagcc tacctagact cagccggctc tccacgcttt gcctgaccct gcttgctcaa
     4741 ctctacgtct ttgtttcgtt ttctgttctg cgccgttaca gatccaagct gtgaccggcg
     4801 cctacgctag gccaccatgg ctgaacaaga tggattgcac gcaggttctc cggccgcttg
     4861 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     4921 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     4981 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     5041 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     5101 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     5161 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     5221 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     5281 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     5341 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     5401 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     5461 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     5521 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     5581 cttctatcgc cttcttgacg agttcttctg agtcgacaat caacctctgg attacaaaat
     5641 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     5701 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     5761 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     5821 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     5881 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     5941 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     6001 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct
     6061 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     6121 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     6181 gatctccctt tgggccgcct ccccgcctgg tacctttaag accaatgact tacaaggcag
     6241 ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc
     6301 aacgaaaata agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag
     6361 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
     6421 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
     6481 gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc
     6541 agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg tttattgcag
     6601 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     6661 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggctct
     6721 agctatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     6781 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     6841 aggaggcttt tttggaggcc tagacttttg cagagacggc ccaaattcgt aatcatggtc
     6901 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     6961 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     7021 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     7081 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     7141 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     7201 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     7261 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     7321 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     7381 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     7441 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     7501 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     7561 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     7621 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     7681 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     7741 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     7801 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     7861 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     7921 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     7981 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     8041 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     8101 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     8161 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     8221 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     8281 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     8341 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     8401 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     8461 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     8521 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     8581 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     8641 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     8701 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     8761 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     8821 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     8881 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     8941 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     9001 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     9061 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     9121 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     9181 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     9241 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     9301 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca
     9361 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     9421 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     9481 ttcccagtca cgacgttgta aaacgacggc cagtgccaag ctg
//
