LOCUS       V043306                 8681 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043306
VERSION     V043306
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     promoter        131..334
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             417..968
                     /label="TNFRSF17(NM_001192)"
                     /note="TNFRSF17(NM_001192)"
                     /gene="TNFRSF17"
     CDS             1014..1724
                     /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"
     misc_feature    1764..1881
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        1930..2440
                     /note="human phosphoglycerate kinase 1 promoter"
                     /label="hPGK promoter"
     CDS             2462..3061
                     /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"
     LTR             3189..3422
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    3494..3615
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      3655..3790
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     promoter        complement(3811..3829)
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     primer_bind     complement(3839..3855)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     rep_origin      3997..4452
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4478..4582
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             4583..5443
                     /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"
     rep_origin      5614..6202
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     protein_bind    6490..6511
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        6526..6556
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6564..6580
                     /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"
     primer_bind     6588..6604
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     promoter        6625..6643
                     /note="promoter for bacteriophage T3 RNA polymerase"
                     /label="T3 promoter"
     promoter        6671..6897
                     /note="Rous sarcoma virus enhancer/promoter"
                     /label="RSV promoter"
     LTR             6898..7078
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /label="5' LTR (truncated)"
     misc_feature    7125..7250
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    7743..7976
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             8161..8205
                     /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"
ORIGIN
        1 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
       61 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      121 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      181 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      241 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      301 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta
      361 gcgctaccgg actcagatct cgagctcaag cttcgaattc tgcagtcgac gccaccatgt
      421 tgcagatggc tgggcagtgc tcccaaaatg aatattttga cagtttgttg catgcttgca
      481 taccttgtca acttcgatgt tcttctaata ctcctcctct aacatgtcag cgttattgta
      541 atgcaagtgt gaccaattca gtgaaaggaa cgaatgcgat tctctggacc tgtttgggac
      601 tgagcttaat aatttctttg gcagttttcg tgctaatgtt tttgctaagg aagataaact
      661 ctgaaccatt aaaggacgag tttaaaaaca caggatcagg tctcctgggc atggctaaca
      721 ttgacctgga aaagagcagg actggtgatg aaattattct tccgagaggc ctcgagtaca
      781 cggtggaaga atgcacctgt gaagactgca tcaagagcaa accgaaggtc gactctgacc
      841 attgctttcc actcccagct atggaggaag gcgcaaccat tcttgtcacc acgaaaacga
      901 atgactattg caagagcctg ccagctgctt tgagtgctac ggagatagag aaatcaattt
      961 ctgctagggg cggtggcggt tctggtggcg gtggctccgg cggtggcggt tctatggtga
     1021 gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca
     1081 tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct
     1141 acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct
     1201 gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg
     1261 acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga
     1321 acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt
     1381 tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga
     1441 agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga
     1501 agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca
     1561 agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca
     1621 agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg
     1681 agggccgcca ctccaccggc ggcatggacg agctgtacaa gtgaattctc gacctcgaga
     1741 caaatggcag tattcatcca caattttaaa agaaaagggg ggattggggg gtacagtgca
     1801 ggggaaagaa tagtagacat aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa
     1861 attacaaaaa ttcaaaattt tcgggtttat tacagggaca gcagagatcc actttggccg
     1921 cggctcgagg gggttggggt tgcgcctttt ccaaggcagc cctgggtttg cgcagggacg
     1981 cggctgctct gggcgtggtt ccgggaaacg cagcggcgcc gaccctgggt ctcgcacatt
     2041 cttcacgtcc gttcgcagcg tcacccggat cttcgccgct acccttgtgg gccccccggc
     2101 gacgcttcct gctccgcccc taagtcggga aggttccttg cggttcgcgg cgtgccggac
     2161 gtgacaaacg gaagccgcac gtctcactag taccctcgca gacggacagc gccagggagc
     2221 aatggcagcg cgccgaccgc gatgggctgt ggccaatagc ggctgctcag cagggcgcgc
     2281 cgagagcagc ggccgggaag gggcggtgcg ggaggcgggg tgtggggcgg tagtgtgggc
     2341 cctgttcctg cccgcgcggt gttccgcatt ctgcaagcct ccggagcgca cgtcggcagt
     2401 cggctccctc gttgaccgaa tcaccgacct ctctccccag ggggatccac cggagcttac
     2461 catgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt
     2521 acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga
     2581 ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga
     2641 catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga
     2701 gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg
     2761 ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga
     2821 gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg
     2881 cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct
     2941 ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc
     3001 cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg
     3061 acgcccgccc cacgacccgc agcgcccgac cgaaaggagc gcacgacccc atgcatcggt
     3121 acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa
     3181 ggggggactg gaagggctaa ttcactccca acgaagacaa gatctgcttt ttgcttgtac
     3241 tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
     3301 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
     3361 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
     3421 cagtagtagt tcatgtcatc ttattattca gtatttataa cttgcaaaga aatgaatatc
     3481 agagagtgag aggaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     3541 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     3601 catcaatgta tcttatcatg tctggctcta gctatcccgc ccctaactcc gcccatcccg
     3661 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     3721 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     3781 tttggaggcc tagggacgta cccaattcgc cctatagtga gtcgtattac gcgcgctcac
     3841 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
     3901 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     3961 cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa
     4021 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     4081 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     4141 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     4201 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     4261 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     4321 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     4381 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     4441 cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     4501 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     4561 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     4621 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     4681 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     4741 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     4801 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     4861 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     4921 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     4981 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     5041 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     5101 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     5161 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     5221 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     5281 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     5341 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     5401 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     5461 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     5521 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     5581 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     5641 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     5701 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     5761 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     5821 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     5881 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     5941 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     6001 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     6061 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     6121 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     6181 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     6241 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     6301 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     6361 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     6421 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     6481 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct
     6541 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta
     6601 tgaccatgat tacgccaagc gcgcaattaa ccctcactaa agggaacaaa agctggagct
     6661 gcaagcttaa tgtagtctta tgcaatactc ttgtagtctt gcaacatggt aacgatgagt
     6721 tagcaacatg ccttacaagg agagaaaaag caccgtgcat gccgattggt ggaagtaagg
     6781 tggtacgatc gtgccttatt aggaaggcaa cagacgggtc tgacatggat tggacgaacc
     6841 actgaattgc cgcattgcag agatattgta tttaagtgcc tagctcgata cataaacggg
     6901 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
     6961 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
     7021 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
     7081 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga
     7141 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa
     7201 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc
     7261 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat
     7321 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg
     7381 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc
     7441 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc
     7501 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa
     7561 acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat
     7621 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
     7681 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
     7741 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
     7801 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
     7861 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
     7921 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
     7981 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
     8041 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
     8101 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
     8161 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
     8221 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
     8281 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
     8341 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
     8401 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atctcgacgg
     8461 tatcgatcac gagactagcc tcgagcggcc gcccccttca ccgagggcct atttcccatg
     8521 attccttcat atttgcatat acgatacaag gctgttagag agataattgg aattaatttg
     8581 actgtaaaca caaagatatt agtacaaaat acgtgacgta gaaagtaata atttcttggg
     8641 tagtttgcag ttttaaaatt atgttttaaa atggactatc a
//
