LOCUS       V042428                 9822 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042428
VERSION     V042428
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             2303..4654
                     /label="Cdh5(NM_009868)"
                     /note="Cdh5(NM_009868)"
                     /gene="Cdh5"
     CDS             4661..4687
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             4691..4717
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             4721..4747
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     promoter        4792..5003
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             5016..5284
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5311..5910
                     /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    5917..6505
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6579..6812
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6884..7005
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      7024..7159
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7197..7213)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7221..7237
                     /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(7245..7275)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7290..7311
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7599..8187)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8358..9218)
                     /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(9219..9323)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9797..9813
                     /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 aattccgcca ccatgcagag gctcacagag ctggccactg ctttgggagc
     2341 cttcctgggc ctcctggcag tggcagcaat ggcaggccct aactttcccc agatagacac
     2401 ccccaacatg ctacctgccc accatcgcca aaagagagac tggatttgga atcaaatgca
     2461 catcgatgaa gagaaaaatg aatcgctgcc ccactatgtg ggaaagatca agtccaacgt
     2521 gaaccgccag aatgctaagt atgtgctcca aggagagttt gctggcaaga tcttcggggt
     2581 cgatgctaac acagggaatg tgcttgccta tgagaggcta gaccgggaga aagtctctga
     2641 gtacttcctt actgccctca ttgtggacaa gaacaccaac aaaaacctgg aacaaccttc
     2701 cagcttcact gtcaaggtgc atgatataaa tgacaattgg cctgtgtttt cgcaccaggt
     2761 attcaacgca tctgtgccag agatgtcagc tatagggacc tctgtcatcc gtgtgacagc
     2821 agtggatgca gatgacccca ctgtggccgg ccacgccact gtcttgtacc aaatcgtgaa
     2881 aggaaatgag tatttcagca ttgataattc tggactcatt ttcacaaaga tcaaaaactt
     2941 ggaccgagag aaacaggctg aatacaagat cgtggtggaa acacaagatg ccctgggtct
     3001 gcggggagag tcaggcacag ccactgtgat gatcagactg gaggacatca atgacaactt
     3061 ccccgtcttt actcaatcca catacacatt ttcggtgcct gaagacatcc gagtgggcaa
     3121 gcccttgggc tttctgactg ttgtggaccc agatgagcct cagaaccgga tgaccaagta
     3181 cagcatcatg cagggcgagt acagggacac cttcaccatt gagacagacc ccaaacgtaa
     3241 cgagggtatc atcaaaccca cgaagtccct ggactatgaa gtcatccagc agtacacatt
     3301 ctacattgag gccacagacc ccactatccg atacgaatac ctgagcagca cttcaggcaa
     3361 aaacaaagcc atggtcacca tcaacgtcct agatgtggat gagccccctg tcttccagcg
     3421 acacttctac cacttcaagc tgccagaaaa ccagaagaaa ccgctgatcg gcactgtggt
     3481 ggccaaagac cctgacaagg ctcagcgcag catcgggtac tccatccgca agaccagtga
     3541 cagaggccaa ttcttccgaa taaccaagca gggaaacatc tataacgaga aagaactgga
     3601 cagagaaacc tacgcctggt ataacctgac tgtggaggcc aacgaactgg attctcgggg
     3661 taaccctgta gggaaagagt ccattgtgca ggtttacatt gaagtcttgg atgagaatga
     3721 caaccctccg gaatttgccc agccctacga acctaaagtg tgtgagaatg ctgcccaggg
     3781 caagctggta gtacagatct ctgcaacaga caaggatgtg gtgccagtaa acccaaagtt
     3841 caagtttgcc ctgaagaacg aggacagcaa cttcaccctc ataaacaacc atgacaacac
     3901 cgccaacatc acggtcaagt atgggcagtt taatcgggag catgccaagt tccactacct
     3961 gcccgtgctc atctcagaca acggggtgcc cagcctgact ggaaccagca cgctaaccgt
     4021 gggtgtgtgc aagtgcaacg agcagggcga gttcaccttc tgtgaggaga tggcagccca
     4081 ggcgggtgtc agcatccagg cgctggtggc catcttcctc tgcatcctca ccatcacagt
     4141 gattaccttg ctgatcatcc tgcggaggcg gatccggaag caggcgcatg ctcatagcaa
     4201 gagtgcgctg gagattcacg agcagttggt cacttacgat gaggagggcg gtggtgagat
     4261 ggacaccacc agctatgatg tgtcagtgct taattctgtg cgcggtggct ccactaagcc
     4321 cctgcgctcc acgatggacg cccggccggc agtgtacacg caggtgcaga agccgccgcg
     4381 cctggcgcct ggactgcacg gagggcccag ggagatggcc accatgatcg acgtgaagaa
     4441 agaagaggca gacaatgacg gcggtggccc cccatacgac acactgcaca tctacggcta
     4501 cgagggcgca gagtccatcg cagagtccct cagttccctg agcaccaatt cctccgactc
     4561 tgacatcgat tatgacttcc tcaacgactg gggacccagg ttcaagatgc tggctgagct
     4621 gtacggctca gatccccagg aggaactcat catcggatcc tacccatacg acgtacctga
     4681 ttatgcaact tatccttatg atgtacctga ttatgctagc tatccttatg atgtcccaga
     4741 ctacgcatag gcggccgcga aggatctgcg atcgctccgg tgcccgtcag tgggcagagc
     4801 gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga acgggtgcct
     4861 agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc
     4921 ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca
     4981 acgggtttgc cgccagaaca cagctgaagc ttcgaggggc tcgcatctct ccttcacgcg
     5041 cccgccgccc tacctgaggc cgccatccac gccggttgag tcgcgttctg ccgcctcccg
     5101 cctgtggtgc ctcctgaact gcgtccgccg tctaggtaag tttaaagctc aggtcgagac
     5161 cgggcctttg tccggcgctc ccttggagcc tacctagact cagccggctc tccacgcttt
     5221 gcctgaccct gcttgctcaa ctctacgtct ttgtttcgtt ttctgttctg cgccgttaca
     5281 gatccaagct gtgaccggcg cctacgctag atgaccgagt acaagcccac ggtgcgcctc
     5341 gccacccgcg acgacgtccc ccgggccgta cgcaccctcg ccgccgcgtt cgccgactac
     5401 cccgccacgc gccacaccgt cgacccggac cgccacatcg agcgggtcac cgagctgcaa
     5461 gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc
     5521 gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag
     5581 atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa
     5641 ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc
     5701 tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg
     5761 gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc
     5821 tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc
     5881 tggtgcatga cccgcaagcc cggtgcctga gtcgacaatc aacctctgga ttacaaaatt
     5941 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     6001 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     6061 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     6121 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     6181 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     6241 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     6301 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg
     6361 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     6421 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     6481 atctcccttt gggccgcctc cccgcctggt acctttaaga ccaatgactt acaaggcagc
     6541 tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca
     6601 acgaaaataa gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc
     6661 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
     6721 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
     6781 acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca
     6841 gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc
     6901 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     6961 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta
     7021 gctatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     7081 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     7141 ggaggctttt ttggaggcct agacttttgc agagacggcc caaattcgta atcatggtca
     7201 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     7261 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     7321 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     7381 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     7441 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     7501 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     7561 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     7621 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     7681 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     7741 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     7801 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     7861 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     7921 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     7981 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     8041 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     8101 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     8161 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     8221 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     8281 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     8341 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     8401 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     8461 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     8521 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     8581 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     8641 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     8701 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     8761 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     8821 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     8881 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     8941 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     9001 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     9061 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     9121 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     9181 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     9241 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     9301 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     9361 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc
     9421 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     9481 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     9541 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
     9601 catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat
     9661 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     9721 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     9781 tcccagtcac gacgttgtaa aacgacggcc agtgccaagc tg
//
