LOCUS       T7opt_in_pCAGGS.        7385 bp DNA     circular SYN 17-JUN-2021
DEFINITION  Human codon-optimized T7 polymerase.
ACCESSION   .
VERSION     .
KEYWORDS    T7opt in pCAGGS.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7385)
  AUTHORS   Yun T, Park A, Hill TE, Pernet O, Beaty SM, Juelich TL, Smith JK, 
            Zhang L, Wang YE, Vigant F, Gao J, Wu P, Lee B, Freiberg AN
  TITLE     Efficient reverse genetics reveals genetic determinants of budding 
            and fusogenic differences between Nipah and Hendra viruses and 
            enables real-time monitoring of viral spread in small animal models 
            of henipavirus infection.
  JOURNAL   J Virol. 2015 Jan 15;89(2):1242-53. doi: 10.1128/JVI.02583-14. Epub 
            2014 Nov 12.
  PUBMED    25392218
REFERENCE   2  (bases 1 to 7385)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 7385)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1128/JVI.02583-14"; journalName: "J Virol"; date: "2015-01-15-
            15"; volume: "89"; issue: "2"; pages: "1242-53"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7385
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        63..366
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        368..645
                     /label=chicken beta-actin promoter
     intron          646..1662
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and
                     rabbit beta-globin"
     primer_bind     1670..1689
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids,
                     forward primer"
     CDS             1723..4371
                     /note="T7 RNA polymerase from Escherichia phage T7.
                     Accession#: P00573"
     primer_bind     complement(4389..4408)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4454..4509
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4508..4527)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also
                     called rb-glob-pA-term-R"
     primer_bind     complement(4870..4886)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4894..4910)
                     /label=lac operator
                     /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-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(4918..4948)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4963..4984)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        5042..5238
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     polyA_signal    5244..5378
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5446..5463)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5617..6205)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6379..7236)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7237..7341)
                     /label=AmpR promoter
     promoter        7369..7385
                     /label=CAG
                     /note="CMV early enhancer fused to modified chicken 
                     beta-actin promoter"
ORIGIN
        1 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
       61 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
      121 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
      181 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
      241 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
      301 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
      361 accatggtcg aggtgagccc cacgttctgc ttcactctcc ccatctcccc cccctcccca
      421 cccccaattt tgtatttatt tattttttaa ttattttgtg cagcgatggg ggcggggggg
      481 gggggggggc gcgcgccrgg sggggsgggg sggggsgrgg ggsggggsgg ggcgaggcgg
      541 agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa agtttccttt tatggcgagg
      601 cggcggcggc ggcggcccta taaaaagcga agcgcgcggc gggcgggagt cgctgcgcgc
      661 tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg
      721 accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattag
      781 cgcttggttt aatgacggct tgtttctttt ctgtggctgc gtgaaagcct tgaggggctc
      841 cgggagggcc ctttgtgcgg ggggagcggc tcggggggtg cgtgcgtgtg tgtgtgcgtg
      901 gggagcgccg cgtgcggctc cgcgctgccc ggcggctgtg agcgctgcgg gcgcggcgcg
      961 gggctttgtg cgctccgcag tgtgcgcgag gggagcgcgg ccgggggcgg tgccccgcgg
     1021 tgcggggggg gctgcgaggg gaacaaaggc tgcgtgcggg gtgtgtgcgt gggggggtga
     1081 gcagggggtg tgggcgcgtc ggtcgggctg caaccccccc tgcacccccc tccccgagtt
     1141 gctgagcacg gcccggcttc gggtgcgggg ctccgtacgg ggcgtggcgc ggggctcgcc
     1201 gtgccgggcg gggggtggcg gcaggtgggg gtgccgggcg gggcggggcc gcctcgggcc
     1261 ggggagggct cgggggaggg gcgcggcggc ccccggagcg ccggcggctg tcgaggcgcg
     1321 gcgagccgca gccattgcct tttatggtaa tcgtgcgaga gggcgcaggg acttcctttg
     1381 tcccaaatct gtgcggagcc gaaatctggg aggcgccgcc gcaccccctc tagcgggcgc
     1441 ggggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg gggagggcct tcgtgcgtcg
     1501 ccgcgccgcc gtccccttct ccctctccag cctcggggct gtccgcgggg ggacggctgc
     1561 cttcgggggg gacggggcag ggcggggttc ggcttctggc gtgtgaccgg cggctctaga
     1621 gcctctgcta accatgttca tgccttcttc tttttcctac agctcctggg caacgtgctg
     1681 gttattgtgc tgtctcatca ttttggcaaa gaattcgcca ccatgaacac catcaatatt
     1741 gccaagaacg acttttctga tatcgagctg gccgctattc cattcaatac actggctgac
     1801 cactacggag agcggctggc ccgcgaacag ctggctctgg agcatgaaag ctatgagatg
     1861 ggagaagccc gattcaggaa gatgtttgag aggcagctga aagctgggga agtggcagac
     1921 aacgcagccg ctaagccact gattaccaca ctgctgccca aaatgatcgc cagaattaat
     1981 gattggttcg aggaagtgaa ggcaaaacga ggaaagaggc ctaccgcctt ccagtttctg
     2041 caggagatca agccagaagc tgtggcatac atcaccatca agactaccct ggcatgcctg
     2101 acaagcgccg acaacacaac tgtgcaggca gtcgcttccg caatcggacg agctattgag
     2161 gacgaagcac gctttgggag aatccgggat ctggaggcca agcacttcaa gaaaaacgtg
     2221 gaggaacagc tgaacaagag ggtggggcat gtctataaga aggccttcat gcaggtggtc
     2281 gaggccgaca tgctgtcaaa gggactgctg ggaggagagg cctggagctc ctggcacaaa
     2341 gaagatagca tccatgtggg agtccgctgc atcgagatgc tgattgaatc tactgggatg
     2401 gtgagtctgc accgacagaa cgccggcgtg gtcggacagg actctgagac aatcgaactg
     2461 gctcccgagt atgccgaagc tattgcaact agagccggag ctctggcagg gatcagtcca
     2521 atgttccagc cctgcgtggt cccccctaag ccttggactg gcatcaccgg gggcggatac
     2581 tgggctaatg gaaggagacc actggcactg gtgcgaacac actctaagaa agccctgatg
     2641 agatacgagg atgtctatat gcccgaagtg tataaggcca tcaacattgc tcagaataca
     2701 gcatggaaaa ttaacaagaa agtgctggcc gtcgctaatg tgatcactaa gtggaaacat
     2761 tgtcccgtgg aggacatccc tgccattgaa cgggaggaac tgcctatgaa gccagaggac
     2821 atcgatatga accctgaagc tctgaccgca tggaagcgcg cagccgctgc agtgtacaga
     2881 aaggataaag cccggaagtc ccggcgcatt tctctggagt tcatgctgga acaggccaac
     2941 aagtttgcta atcacaaagc aatctggttc ccctacaaca tggactggcg cggacgagtc
     3001 tatgccgtga gcatgttcaa ccctcagggg aatgatatga caaagggcct gctgactctg
     3061 gctaagggga aaccaattgg gaaggagggc tactattggc tgaaaatcca cggggccaat
     3121 tgcgctggcg tcgacaaggt gccattcccc gagaggatca agttcatcga ggaaaaccat
     3181 gaaaatatta tggcatgtgc caagtctccc ctggagaaca catggtgggc cgaacaggat
     3241 agtcctttct gctttctggc cttctgtttt gagtacgctg gagtgcagca ccatgggctg
     3301 agttataatt gctccctgcc actggccttt gacggctctt gtagtggaat ccagcacttc
     3361 tccgcaatgc tgagggatga ggtcggagga agagcagtga acctgctgcc atctgagaca
     3421 gtgcaggaca tctacggcat tgtcgccaag aaagtgaatg agatcctgca ggctgacgca
     3481 attaacggga ctgataatga ggtggtcacc gtcacagatg aaaacactgg cgagatcagc
     3541 gaaaaggtga aactgggaac caaggccctg gctggacagt ggctggcata cggagtcacc
     3601 cgctcagtga caaagcgaag cgtgatgacc ctggcttatg gcagcaaaga gttcggcttc
     3661 aggcagcagg tgctggaaga caccatccag ccagccattg attccggaaa ggggctgatg
     3721 tttacacagc ccaaccaggc cgctggctac atggccaagc tgatctggga gtcagtgagc
     3781 gtcacagtgg tcgcagccgt ggaagctatg aattggctga agtccgctgc aaaactgctg
     3841 gccgctgagg tgaaggacaa gaaaactggc gaaattctga ggaaaagatg cgccgtccac
     3901 tgggtgaccc ctgatggatt cccagtgtgg caggagtata agaaacccat ccagaccaga
     3961 ctgaacctga tgttcctggg ccagtttcgg ctgcagccta caatcaacac taataaggac
     4021 agtgagattg atgctcataa acaggaatca gggattgcac ctaattttgt gcacagccag
     4081 gacggctccc atctgcggaa gactgtggtc tgggctcacg agaaatacgg catcgaatcc
     4141 ttcgcactga ttcatgactc ttttggaacc atcccagccg atgcagccaa cctgttcaag
     4201 gctgtccgcg agactatggt ggacacctac gaaagttgtg atgtgctggc cgacttctat
     4261 gatcagtttg ctgaccagct gcacgagtca cagctggata agatgcccgc actgcctgcc
     4321 aaaggcaacc tgaatctgag agacatcctg gagtccgatt tcgcatttgc ctgactcgag
     4381 ctagcagatc tttttccctc tgccaaaaat tatggggaca tcatgaagcc ccttgagcat
     4441 ctgacttctg gctaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg
     4501 tgtctctcac tcggaaggac atatgggagg gcaaatcatt taaaacatca gaatgagtat
     4561 ttggtttaga gtttggcaac atatgcccat atgctggctg ccatgaacaa aggttggcta
     4621 taaagaggtc atcagtatat gaaacagccc cctgctgtcc attccttatt ccatagaaaa
     4681 gccttgactt gaggttagat tttttttata ttttgttttg tgttattttt ttctttaaca
     4741 tccctaaaat tttccttaca tgttttacta gccagatttt tcctcctctc ctgactactc
     4801 ccagtcatag ctgtccctct tctcttatgg agatccctcg acctgcagcc caagcttggc
     4861 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     4921 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     4981 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagcggat
     5041 ccgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     5101 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     5161 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     5221 ggcctaggct tttgcaaaaa gctaacttgt ttattgcagc ttataatggt tacaaataaa
     5281 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     5341 tgtccaaact catcaatgta tcttatcatg tctggatccg ctgcattaat gaatcggcca
     5401 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     5461 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     5521 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     5581 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     5641 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     5701 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     5761 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     5821 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     5881 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     5941 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     6001 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     6061 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     6121 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     6181 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     6241 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     6301 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     6361 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     6421 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     6481 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     6541 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     6601 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     6661 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     6721 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     6781 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     6841 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     6901 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     6961 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     7021 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     7081 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     7141 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     7201 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     7261 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     7321 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgggt cgacattgat
     7381 tattg
//
