LOCUS       V045449                 9893 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045449
VERSION     V045449
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..4074
                     /label="Canx(NM_007597)"
                     /note="Canx(NM_007597)"
                     /gene="Canx"
     CDS             4111..4821
                     /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"
     promoter        4863..5074
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             5087..5355
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5382..5981
                     /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    5988..6576
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6650..6883
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6955..7076
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      7095..7230
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7268..7284)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7292..7308
                     /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(7316..7346)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7361..7382
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7670..8258)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8429..9289)
                     /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(9290..9394)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9868..9884
                     /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 catggaaggg aagtggttac tgtgtttgct gctggtcctt
     2341 ggaactgcag ctgttgaggc tcatgatgga catgatgatg acgcgattga tattgaagat
     2401 gatcttgatg atgttattga agaggtagaa gattcaaaat ctaaatcaga tgccagcact
     2461 cctccatctc caaaggtcac ctacaaagct ccagttccaa caggggaggt ttattttgct
     2521 gactcctttg acagagggtc tctgtcaggg tggattttat ctaaagccaa aaaagatgac
     2581 actgatgatg aaattgccaa atatgatgga aagtgggaag tagatgagat gaaggaaaca
     2641 aagcttccag gggataaagg acttgtactg atgtctcggg ccaagcatca tgccatctct
     2701 gctaaactga ataagccctt cctgtttgat accaagcctc tcattgttca gtatgaggtt
     2761 aattttcaga atggaataga atgtggtggt gcctatgtga agctgctttc caagacggca
     2821 gagctcagcc tggatcaatt ccacgacaag actccctata ctattatgtt tggtccagat
     2881 aagtgtggag aggactacaa actgcatttc atctttcgac acaaaaatcc caagacaggt
     2941 gtatatgaag aaaaacatgc taagaggcca gatgcagatc tgaagaccta tttcactgac
     3001 aagaaaacgc atctttatac attaatcttg aatccagaca atagttttga aatattagtt
     3061 gaccagtctg ttgtgaacag tggaaatctg ctaaatgaca tgactcctcc tgtaaaccct
     3121 tcacgtgaaa ttgaagaccc agaagaccgg aagcctgaag attgggatga aaggcccaaa
     3181 atagcagatc cagatgctgt caagccagat gactgggatg aagacgcccc ttctaagatc
     3241 ccagatgaag aggccaccaa gcctgaaggc tggctagacg acgaacctga gtatattcca
     3301 gaccctgatg cagagaagcc agaggattgg gatgaggata tggacggaga atgggaggct
     3361 cctcagattg ccaaccccaa gtgtgagtca gcccctgggt gtggtgtctg gcagcgacct
     3421 atgattgaca accccaatta taagggcaaa tggaagcctc caatgattga caaccctaac
     3481 taccagggaa tctggaaacc aaggaaaata ccaaatccag atttctttga agacctagaa
     3541 ccttttaaga tgactccttt cagtgctatt ggtttggagc tctggtccat gacatccgac
     3601 atcttttttg acaactttat cattagtggt gaccgaagag tagttgatga ttgggccaat
     3661 gatgggtggg gcctgaagaa agctgctgat ggggctgctg agccaggtgt agtgctgcag
     3721 atgctggagg cagctgaaga gcgtccatgg ctttgggtgg tctacatttt gactgtagct
     3781 ttgccagtgt tccttgtgat cctcttctgc tgttctggaa agaaacagtc caatgctatg
     3841 gagtacaaga agacggatgc tccccagcca gatgtgaagg atgaagaagg gaaggaagaa
     3901 gagaagaaca agagggatga agaggaagaa gaggagaagc ttgaagagaa acagaagagt
     3961 gatgctgaag aagatggtgt tactggcagt caagatgagg aagatagcaa gcctaaagca
     4021 gaggaggatg aaattttgaa cagatcgcca agaaacagaa agccacgaag agagggatcc
     4081 ggtggaggcg gatcaggagg cggtggatct atggtgagca agggcgagga ggataacatg
     4141 gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt gaacggccac
     4201 gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag
     4261 ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc
     4321 atgtacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttgaagctg
     4381 tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg
     4441 accgtgaccc aggactcctc cctgcaggac ggcgagttca tctacaaggt gaagctgcgc
     4501 ggcaccaact tcccctccga cggccccgta atgcagaaga agaccatggg ctgggaggcc
     4561 tcctccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa gcagaggctg
     4621 aagctgaagg acggcggcca ctacgacgct gaggtcaaga ccacctacaa ggccaagaag
     4681 cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt tggacatcac ctcccacaac
     4741 gaggactaca ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc
     4801 atggacgagc tgtacaagta ggcggccgcg aaggatctgc gatcgctccg gtgcccgtca
     4861 gtgggcagag cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg
     4921 aacgggtgcc tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct
     4981 ccgccttttt cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt
     5041 tctttttcgc aacgggtttg ccgccagaac acagctgaag cttcgagggg ctcgcatctc
     5101 tccttcacgc gcccgccgcc ctacctgagg ccgccatcca cgccggttga gtcgcgttct
     5161 gccgcctccc gcctgtggtg cctcctgaac tgcgtccgcc gtctaggtaa gtttaaagct
     5221 caggtcgaga ccgggccttt gtccggcgct cccttggagc ctacctagac tcagccggct
     5281 ctccacgctt tgcctgaccc tgcttgctca actctacgtc tttgtttcgt tttctgttct
     5341 gcgccgttac agatccaagc tgtgaccggc gcctacgcta gatgaccgag tacaagccca
     5401 cggtgcgcct cgccacccgc gacgacgtcc cccgggccgt acgcaccctc gccgccgcgt
     5461 tcgccgacta ccccgccacg cgccacaccg tcgacccgga ccgccacatc gagcgggtca
     5521 ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg
     5581 cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg
     5641 tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc
     5701 aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca
     5761 ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg
     5821 gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca
     5881 acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag
     5941 gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg agtcgacaat caacctctgg
     6001 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     6061 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     6121 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
     6181 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
     6241 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
     6301 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     6361 attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca
     6421 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
     6481 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
     6541 agacgagtcg gatctccctt tgggccgcct ccccgcctgg tacctttaag accaatgact
     6601 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta
     6661 attcactccc aacgaaaata agatctgctt tttgcttgta ctgggtctct ctggttagac
     6721 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
     6781 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
     6841 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat
     6901 cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg
     6961 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     7021 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     7081 gtctggctct agctatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     7141 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     7201 agaagtagtg aggaggcttt tttggaggcc tagacttttg cagagacggc ccaaattcgt
     7261 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     7321 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     7381 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     7441 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     7501 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     7561 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     7621 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     7681 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     7741 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     7801 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     7861 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     7921 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     7981 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     8041 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     8101 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     8161 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     8221 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     8281 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     8341 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     8401 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     8461 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     8521 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     8581 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     8641 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     8701 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     8761 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     8821 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     8881 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     8941 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     9001 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     9061 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     9121 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     9181 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     9241 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     9301 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     9361 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     9421 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     9481 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     9541 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     9601 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
     9661 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     9721 tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     9781 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
     9841 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgccaag ctg
//
