LOCUS       V034463                 8902 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034463
VERSION     V034463
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..3531
                     /label="LAMP2(NM_002294)"
                     /note="LAMP2(NM_002294)"
                     /gene="LAMP2"
     CDS             3538..3627
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     promoter        3671..3882
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3895..4163
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4196..4990
                     /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    4997..5585
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5659..5892
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5964..6085
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6104..6239
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6277..6293)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6301..6317
                     /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(6325..6355)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6370..6391
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6679..7267)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7438..8298)
                     /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(8299..8403)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8877..8893
                     /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 catggtgtgc ttccgcctct tcccggttcc gggctcaggg
     2341 ctcgttctgg tctgcctagt cctgggagct gtgcggtctt atgcattgga acttaatttg
     2401 acagattcag aaaatgccac ttgcctttat gcaaaatggc agatgaattt cacagtacgc
     2461 tatgaaacta caaataaaac ttataaaact gtaaccattt cagaccatgg cactgtgaca
     2521 tataatggaa gcatttgtgg ggatgatcag aatggtccca aaatagcagt gcagttcgga
     2581 cctggctttt cctggattgc gaattttacc aaggcagcat ctacttattc aattgacagc
     2641 gtctcatttt cctacaacac tggtgataac acaacatttc ctgatgctga agataaagga
     2701 attcttactg ttgatgaact tttggccatc agaattccat tgaatgacct ttttagatgc
     2761 aatagtttat caactttgga aaagaatgat gttgtccaac actactggga tgttcttgta
     2821 caagcttttg tccaaaatgg cacagtgagc acaaatgagt tcctgtgtga taaagacaaa
     2881 acttcaacag tggcacccac catacacacc actgtgccat ctcctactac aacacctact
     2941 ccaaaggaaa aaccagaagc tggaacctat tcagttaata atggcaatga tacttgtctg
     3001 ctggctacca tggggctgca gctgaacatc actcaggata aggttgcttc agttattaac
     3061 atcaacccca atacaactca ctccacaggc agctgccgtt ctcacactgc tctacttaga
     3121 ctcaatagca gcaccattaa gtatctagac tttgtctttg ctgtgaaaaa tgaaaaccga
     3181 ttttatctga aggaagtgaa catcagcatg tatttggtta atggctccgt tttcagcatt
     3241 gcaaataaca atctcagcta ctgggatgcc cccctgggaa gttcttatat gtgcaacaaa
     3301 gagcagactg tttcagtgtc tggagcattt cagataaata cctttgatct aagggttcag
     3361 cctttcaatg tgacacaagg aaagtattct acagctcaag actgcagtgc agatgacgac
     3421 aacttccttg tgcccatagc ggtgggagct gccttggcag gagtacttat tctagtgttg
     3481 ctggcttatt ttattggtct caagcaccat catgctggat atgagcaatt tggatccgaa
     3541 caaaaactaa tatcagagga agatttggaa cagaaactaa tcagtgaaga agatttagag
     3601 caaaagttaa tttccgagga ggacttatag cggccgcgaa ggatctgcga tcgctccggt
     3661 gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc
     3721 ggcaattgaa cgggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg
     3781 tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc
     3841 gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac agctgaagct tcgaggggct
     3901 cgcatctctc cttcacgcgc ccgccgccct acctgaggcc gccatccacg ccggttgagt
     3961 cgcgttctgc cgcctcccgc ctgtggtgcc tcctgaactg cgtccgccgt ctaggtaagt
     4021 ttaaagctca ggtcgagacc gggcctttgt ccggcgctcc cttggagcct acctagactc
     4081 agccggctct ccacgctttg cctgaccctg cttgctcaac tctacgtctt tgtttcgttt
     4141 tctgttctgc gccgttacag atccaagctg tgaccggcgc ctacgctagg ccaccatggc
     4201 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     4261 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     4321 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
     4381 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     4441 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     4501 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     4561 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     4621 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     4681 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
     4741 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     4801 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     4861 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     4921 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     4981 gttcttctga gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     5041 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     5101 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     5161 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     5221 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     5281 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     5341 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
     5401 ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     5461 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     5521 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     5581 cccgcctggt acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt
     5641 taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaaaataa gatctgcttt
     5701 ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
     5761 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
     5821 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
     5881 aaatctctag cagtagtagt tcatgtcatc ttattattca gtatttataa cttgcaaaga
     5941 aatgaatatc agagagtgag aggaacttgt ttattgcagc ttataatggt tacaaataaa
     6001 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     6061 tgtccaaact catcaatgta tcttatcatg tctggctcta gctatcccgc ccctaactcc
     6121 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     6181 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     6241 agacttttgc agagacggcc caaattcgta atcatggtca tagctgtttc ctgtgtgaaa
     6301 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     6361 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     6421 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     6481 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     6541 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     6601 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     6661 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     6721 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     6781 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     6841 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     6901 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     6961 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     7021 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     7081 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     7141 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     7201 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     7261 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     7321 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     7381 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     7441 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     7501 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     7561 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     7621 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     7681 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     7741 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     7801 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     7861 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     7921 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     7981 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     8041 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     8101 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     8161 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     8221 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     8281 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     8341 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     8401 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     8461 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     8521 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     8581 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     8641 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
     8701 gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa
     8761 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     8821 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
     8881 aacgacggcc agtgccaagc tg
//
