LOCUS       V045332                 9586 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045332
VERSION     V045332
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..4440
                     /label="Lrp10(NM_022993)"
                     /note="Lrp10(NM_022993)"
                     /gene="Lrp10"
     CDS             4447..4512
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4556..4767
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4780..5048
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5075..5674
                     /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    5681..6269
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6343..6576
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6648..6769
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6788..6923
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6961..6977)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6985..7001
                     /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(7009..7039)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7054..7075
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7363..7951)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8122..8982)
                     /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(8983..9087)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9561..9577
                     /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 catgctatcg gccctccctc ttcttttcct cctcctcgga
     2341 ggtgctctgg cccgcccaga ccggatcact ttcccacgtt ctgcttgtga ggcgccccca
     2401 gctgtgctct cagaagtaca aggcacttta cagaggcctc taggccggga cagccgaagc
     2461 tcccctgcca actgcacctg ggttatcctg ggcagcaagg atcagacagt aactgtcagg
     2521 ttccagaagc tgcatctggc ctgtggctca gagcatttaa tcctgcactc ccctctacag
     2581 ccaccaatct ccctgtgtga ggccccttct ggccctttgc agctaccagg gggcaatgtc
     2641 accattacat acagctatgc tggggccaga gcacccatgg gccagggctt cttgctgact
     2701 tacagtcaag attggctact gtgcctgcaa gaagaattcc agtgcctgaa ccaccgctgc
     2761 attccagctg ctcagcgctg tgatgggatt gatgcctgcg gggatggctc agacgaggca
     2821 ggctgcagct cagatccatt ccctaacctg aacccagccc ccgcaccaac tctggcctgc
     2881 aatctcacct tggaggactt ttatggggtc ttttcttccc ctggatattc acacctggcc
     2941 tcagtctccc acccccagtc ctgcctgtgg ctgctggacc cccatgatgg ccggaggctg
     3001 gcagtgcgct tcacagccct ggacttgagt tacggagatg cagtgcatgt gtatgatggt
     3061 gctggacccc ccgagacccc tcgactgcta cgtagcctca cccacttcag caatggcaag
     3121 gctgtcactg tggagaccct gtctggtcag gctgttgtgt cctaccacac agttgcttgg
     3181 agcagtggcc ggggctttaa tgctacctac catgtccggg gctactgttt accttgggac
     3241 agaccctgtg gcttgggctc tggcctgggg gctagtgaga acctaggtga gcgttgctat
     3301 agcgaggcac agcgctgtga tggctcatgg gactgtgccg atggcacaga tgaggagggt
     3361 tgccctggct gcccaccagg gcacttcccc tgtggagctg caggcacccc tggtgccaca
     3421 gcctgctacc tgcctgctga ccgctgcaac taccagacgt tctgcgccga tggagcggat
     3481 gagaggcgct gccggcattg ccagcccggc aacttccggt gccgggatga gaagtgtgtg
     3541 tatgagacat gggtgtgtga tgggcagcca gactgtactg acggcagtga tgagtgggac
     3601 tgctcctacg ccctgccccg aaaagtcatc acagcagcag tcattggcag cctggtgtgt
     3661 ggcctgttgc tggtcatcgc tctcggctgc acctgcaaac tctatgccat ccgcacccag
     3721 gaatacagca tctttgcccc gctctcccgg atggaggctg agattgtgca gcaacaggca
     3781 cccccttcct atgggcagct cattgcccag ggtgccatcc cgcctgtgga agacttcccc
     3841 acagagaacc ctaacgataa ctctgtgctg ggaaacctac gttctctgct tcagatctta
     3901 cgccaggata tgactccagg tggtacttca gggggccgcc gtcgccagcg tggacgctcc
     3961 atccgccgtc tggttcgccg tctccgtcgt tggggcctgc ttcctcgaac taatacccca
     4021 gctcgggccc ctgagaccag atcccaggtc acaccctctg ttccctctga ggccctggat
     4081 gacagcacag gccaagcctg tgagggtggg gcagtagggg ggcaagatgg ggagcaggct
     4141 cctccactgc ccatcaagac ccccatccca accccaagca cacttcctgc ccttgctact
     4201 gtctctgagc ccccagggcc actaccctca gtgcctgtag aatcatcact gttgtctgga
     4261 gttgtccagg tcctacgagg ccgcctccta cccagcctgt ggtccccagg ccccacttgg
     4321 acccaaactg gaactcacac aacagtcctg tccccagagg atgaggacga tgtattgtta
     4381 ttgccactgg ctgagccaga agtctgggtg gtggaggcag aggatgaacc attgcttgcc
     4441 ggatccgact acaaagacca tgacggtgat tataaagatc atgacatcga ctacaaggat
     4501 gacgatgaca agtagcggcc gcgaaggatc tgcgatcgct ccggtgcccg tcagtgggca
     4561 gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaacgggt
     4621 gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt
     4681 tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt
     4741 cgcaacgggt ttgccgccag aacacagctg aagcttcgag gggctcgcat ctctccttca
     4801 cgcgcccgcc gccctacctg aggccgccat ccacgccggt tgagtcgcgt tctgccgcct
     4861 cccgcctgtg gtgcctcctg aactgcgtcc gccgtctagg taagtttaaa gctcaggtcg
     4921 agaccgggcc tttgtccggc gctcccttgg agcctaccta gactcagccg gctctccacg
     4981 ctttgcctga ccctgcttgc tcaactctac gtctttgttt cgttttctgt tctgcgccgt
     5041 tacagatcca agctgtgacc ggcgcctacg ctagatgacc gagtacaagc ccacggtgcg
     5101 cctcgccacc cgcgacgacg tcccccgggc cgtacgcacc ctcgccgccg cgttcgccga
     5161 ctaccccgcc acgcgccaca ccgtcgaccc ggaccgccac atcgagcggg tcaccgagct
     5221 gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga
     5281 cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc
     5341 cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat
     5401 ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg
     5461 cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga
     5521 ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc
     5581 cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg
     5641 cacctggtgc atgacccgca agcccggtgc ctgagtcgac aatcaacctc tggattacaa
     5701 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     5761 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     5821 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     5881 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     5941 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     6001 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     6061 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat
     6121 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     6181 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     6241 tcggatctcc ctttgggccg cctccccgcc tggtaccttt aagaccaatg acttacaagg
     6301 cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact
     6361 cccaacgaaa ataagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct
     6421 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     6481 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     6541 tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta
     6601 ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg
     6661 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     6721 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc
     6781 tctagctatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     6841 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     6901 gtgaggaggc ttttttggag gcctagactt ttgcagagac ggcccaaatt cgtaatcatg
     6961 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     7021 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     7081 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     7141 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     7201 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     7261 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     7321 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     7381 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     7441 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     7501 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     7561 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     7621 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     7681 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     7741 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     7801 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     7861 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     7921 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     7981 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     8041 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     8101 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     8161 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     8221 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     8281 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     8341 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     8401 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     8461 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     8521 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     8581 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     8641 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     8701 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     8761 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     8821 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     8881 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     8941 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     9001 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     9061 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     9121 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     9181 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     9241 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     9301 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     9361 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     9421 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     9481 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     9541 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctg
//
