LOCUS       V045045                 9458 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045045
VERSION     V045045
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..3009
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /transl_table=1
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
                     /label="mCherry"
     CDS             3040..4290
                     /label="CALR(NM_004343)"
                     /note="CALR(NM_004343)"
                     /gene="CALR"
     CDS             4297..4323
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             4327..4353
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             4357..4383
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     promoter        4428..4639
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4652..4920
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4947..5546
                     /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    5553..6141
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6215..6448
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6520..6641
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6660..6795
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6833..6849)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6857..6873
                     /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(6881..6911)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6926..6947
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7235..7823)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7994..8854)
                     /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(8855..8959)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9433..9449
                     /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 catggtgagc aagggcgagg aggataacat ggccatcatc
     2341 aaggagttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca cgagttcgag
     2401 atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg
     2461 accaagggtg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt catgtacggc
     2521 tccaaggcct acgtgaagca ccccgccgac atccccgact acttgaagct gtccttcccc
     2581 gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc
     2641 caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggcaccaac
     2701 ttcccctccg acggccccgt aatgcagaag aagaccatgg gctgggaggc ctcctccgag
     2761 cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agcagaggct gaagctgaag
     2821 gacggcggcc actacgacgc tgaggtcaag accacctaca aggccaagaa gcccgtgcag
     2881 ctgcccggcg cctacaacgt caacatcaag ttggacatca cctcccacaa cgaggactac
     2941 accatcgtgg aacagtacga acgcgccgag ggccgccact ccaccggcgg catggacgag
     3001 ctgtacaagg gaggtggagg tagtggagga ggcggatcca tgctgctatc cgtgccgctg
     3061 ctgctcggcc tcctcggcct ggccgtcgcc gagcctgccg tctacttcaa ggagcagttt
     3121 ctggacggag acgggtggac ttcccgctgg atcgaatcca aacacaagtc agattttggc
     3181 aaattcgttc tcagttccgg caagttctac ggtgacgagg agaaagataa aggtttgcag
     3241 acaagccagg atgcacgctt ttatgctctg tcggccagtt tcgagccttt cagcaacaaa
     3301 ggccagacgc tggtggtgca gttcacggtg aaacatgagc agaacatcga ctgtgggggc
     3361 ggctatgtga agctgtttcc taatagtttg gaccagacag acatgcacgg agactcagaa
     3421 tacaacatca tgtttggtcc cgacatctgt ggccctggca ccaagaaggt tcatgtcatc
     3481 ttcaactaca agggcaagaa cgtgctgatc aacaaggaca tccgttgcaa ggatgatgag
     3541 tttacacacc tgtacacact gattgtgcgg ccagacaaca cctatgaggt gaagattgac
     3601 aacagccagg tggagtccgg ctccttggaa gacgattggg acttcctgcc acccaagaag
     3661 ataaaggatc ctgatgcttc aaaaccggaa gactgggatg agcgggccaa gatcgatgat
     3721 cccacagact ccaagcctga ggactgggac aagcccgagc atatccctga ccctgatgct
     3781 aagaagcccg aggactggga tgaagagatg gacggagagt gggaaccccc agtgattcag
     3841 aaccctgagt acaagggtga gtggaagccc cggcagatcg acaacccaga ttacaagggc
     3901 acttggatcc acccagaaat tgacaacccc gagtattctc ccgatcccag tatctatgcc
     3961 tatgataact ttggcgtgct gggcctggac ctctggcagg tcaagtctgg caccatcttt
     4021 gacaacttcc tcatcaccaa cgatgaggca tacgctgagg agtttggcaa cgagacgtgg
     4081 ggcgtaacaa aggcagcaga gaaacaaatg aaggacaaac aggacgagga gcagaggctt
     4141 aaggaggagg aagaagacaa gaaacgcaaa gaggaggagg aggcagagga caaggaggat
     4201 gatgaggaca aagatgagga tgaggaggat gaggaggaca aggaggaaga tgaggaggaa
     4261 gatgtccccg gccaggccaa ggacgagctg ctcgagtacc catacgacgt acctgattat
     4321 gcaacttatc cttatgatgt acctgattat gctagctatc cttatgatgt cccagactac
     4381 gcataggcgg ccgcgaagga tctgcgatcg ctccggtgcc cgtcagtggg cagagcgcac
     4441 atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgaacgg gtgcctagag
     4501 aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga
     4561 gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg
     4621 gtttgccgcc agaacacagc tgaagcttcg aggggctcgc atctctcctt cacgcgcccg
     4681 ccgccctacc tgaggccgcc atccacgccg gttgagtcgc gttctgccgc ctcccgcctg
     4741 tggtgcctcc tgaactgcgt ccgccgtcta ggtaagttta aagctcaggt cgagaccggg
     4801 cctttgtccg gcgctccctt ggagcctacc tagactcagc cggctctcca cgctttgcct
     4861 gaccctgctt gctcaactct acgtctttgt ttcgttttct gttctgcgcc gttacagatc
     4921 caagctgtga ccggcgccta cgctagatga ccgagtacaa gcccacggtg cgcctcgcca
     4981 cccgcgacga cgtcccccgg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg
     5041 ccacgcgcca caccgtcgac ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac
     5101 tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg
     5161 cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg
     5221 gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc
     5281 tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc
     5341 ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg
     5401 agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg
     5461 agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt
     5521 gcatgacccg caagcccggt gcctgagtcg acaatcaacc tctggattac aaaatttgtg
     5581 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     5641 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     5701 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     5761 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     5821 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     5881 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     5941 cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg
     6001 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     6061 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     6121 ccctttgggc cgcctccccg cctggtacct ttaagaccaa tgacttacaa ggcagctgta
     6181 gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga
     6241 aaataagatc tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg
     6301 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
     6361 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
     6421 ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat
     6481 ttataacttg caaagaaatg aatatcagag agtgagagga acttgtttat tgcagcttat
     6541 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     6601 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gctctagcta
     6661 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     6721 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     6781 gcttttttgg aggcctagac ttttgcagag acggcccaaa ttcgtaatca tggtcatagc
     6841 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     6901 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     6961 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     7021 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     7081 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     7141 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     7201 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     7261 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     7321 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     7381 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     7441 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     7501 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     7561 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     7621 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     7681 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     7741 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     7801 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     7861 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     7921 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     7981 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     8041 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     8101 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     8161 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     8221 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     8281 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     8341 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     8401 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     8461 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     8521 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     8581 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     8641 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     8701 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     8761 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     8821 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     8881 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     8941 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     9001 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc
     9061 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     9121 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     9181 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     9241 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc
     9301 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     9361 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
     9421 agtcacgacg ttgtaaaacg acggccagtg ccaagctg
//
