LOCUS       Exported                7381 bp ds-DNA     circular SYN 12-MAY-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 7381)
  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 7381)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7381
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        368..645
                     /label=chicken beta-actin promoter
     intron          647..1664
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1588..1610
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1672..1691
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(4385..4404)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4450..4505
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4504..4523)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4866..4882)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4866..4882)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4879..4901)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4890..4906
                     /label=lac operator
                     /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-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(4914..4944)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4959..4980
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5039..5234
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5085..5220
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5147..5166
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    5240..5374
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5277..5296)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5331..5350
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5442..5459)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5613..6201)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5693..5712)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6372..7232)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6995..7014
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7233..7337)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        7368..366
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
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 gcgcgccagg cggggcgggg cggggcgagg ggcggggcgg ggcgaggcgg
      541 agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa agtttccttt tatggcgagg
      601 cggcggcggc ggcggcccta taaaaagcga agcgcgcggc gggcggggag tcgctgcgac
      661 gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc cggctctgac
      721 tgaccgcgtt actcccacag gtgagcgggc gggacggccc ttctcctccg ggctgtaatt
      781 agcgcttggt ttaatgacgg cttgtttctt ttctgtggct gcgtgaaagc cttgaggggc
      841 tccgggaggg ccctttgtgc ggggggagcg gctcgggggg tgcgtgcgtg tgtgtgtgcg
      901 tggggagcgc cgcgtgcggc tccgcgctgc ccggcggctg tgagcgctgc gggcgcggcg
      961 cggggctttg tgcgctccgc agtgtgcgcg aggggagcgc ggccgggggc ggtgccccgc
     1021 ggtgcggggg gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc gtgggggggt
     1081 gagcaggggg tgtgggcgcg tcggtcgggc tgcaaccccc cctgcacccc cctccccgag
     1141 ttgctgagca cggcccggct tcgggtgcgg ggctccgtac ggggcgtggc gcggggctcg
     1201 ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg ccgcctcggg
     1261 ccggggaggg ctcgggggag gggcgcggcg gcccccggag cgccggcggc tgtcgaggcg
     1321 cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag ggacttcctt
     1381 tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc tctagcgggc
     1441 gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc cttcgtgcgt
     1501 cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg ggggacggct
     1561 gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc ggcggctcta
     1621 gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg ggcaacgtgc
     1681 tggttattgt gctgtctcat cattttggca aagaattcgc caccatgaac accatcaata
     1741 ttgccaagaa cgacttttct gatatcgagc tggccgctat tccattcaat acactggctg
     1801 accactacgg agagcggctg gcccgcgaac agctggctct ggagcatgaa agctatgaga
     1861 tgggagaagc ccgattcagg aagatgtttg agaggcagct gaaagctggg gaagtggcag
     1921 acaacgcagc cgctaagcca ctgattacca cactgctgcc caaaatgatc gccagaatta
     1981 atgattggtt cgaggaagtg aaggcaaaac gaggaaagag gcctaccgcc ttccagtttc
     2041 tgcaggagat caagccagaa gctgtggcat acatcaccat caagactacc ctggcatgcc
     2101 tgacaagcgc cgacaacaca actgtgcagg cagtcgcttc cgcaatcgga cgagctattg
     2161 aggacgaagc acgctttggg agaatccggg atctggaggc caagcacttc aagaaaaacg
     2221 tggaggaaca gctgaacaag agggtggggc atgtctataa gaaggccttc atgcaggtgg
     2281 tcgaggccga catgctgtca aagggactgc tgggaggaga ggcctggagc tcctggcaca
     2341 aagaagatag catccatgtg ggagtccgct gcatcgagat gctgattgaa tctactggga
     2401 tggtgagtct gcaccgacag aacgccggcg tggtcggaca ggactctgag acaatcgaac
     2461 tggctcccga gtatgccgaa gctattgcaa ctagagccgg agctctggca gggatcagtc
     2521 caatgttcca gccctgcgtg gtccccccta agccttggac tggcatcacc gggggcggat
     2581 actgggctaa tggaaggaga ccactggcac tggtgcgaac acactctaag aaagccctga
     2641 tgagatacga ggatgtctat atgcccgaag tgtataaggc catcaacatt gctcagaata
     2701 cagcatggaa aattaacaag aaagtgctgg ccgtcgctaa tgtgatcact aagtggaaac
     2761 attgtcccgt ggaggacatc cctgccattg aacgggagga actgcctatg aagccagagg
     2821 acatcgatat gaaccctgaa gctctgaccg catggaagcg cgcagccgct gcagtgtaca
     2881 gaaaggataa agcccggaag tcccggcgca tttctctgga gttcatgctg gaacaggcca
     2941 acaagtttgc taatcacaaa gcaatctggt tcccctacaa catggactgg cgcggacgag
     3001 tctatgccgt gagcatgttc aaccctcagg ggaatgatat gacaaagggc ctgctgactc
     3061 tggctaaggg gaaaccaatt gggaaggagg gctactattg gctgaaaatc cacggggcca
     3121 attgcgctgg cgtcgacaag gtgccattcc ccgagaggat caagttcatc gaggaaaacc
     3181 atgaaaatat tatggcatgt gccaagtctc ccctggagaa cacatggtgg gccgaacagg
     3241 atagtccttt ctgctttctg gccttctgtt ttgagtacgc tggagtgcag caccatgggc
     3301 tgagttataa ttgctccctg ccactggcct ttgacggctc ttgtagtgga atccagcact
     3361 tctccgcaat gctgagggat gaggtcggag gaagagcagt gaacctgctg ccatctgaga
     3421 cagtgcagga catctacggc attgtcgcca agaaagtgaa tgagatcctg caggctgacg
     3481 caattaacgg gactgataat gaggtggtca ccgtcacaga tgaaaacact ggcgagatca
     3541 gcgaaaaggt gaaactggga accaaggccc tggctggaca gtggctggca tacggagtca
     3601 cccgctcagt gacaaagcga agcgtgatga ccctggctta tggcagcaaa gagttcggct
     3661 tcaggcagca ggtgctggaa gacaccatcc agccagccat tgattccgga aaggggctga
     3721 tgtttacaca gcccaaccag gccgctggct acatggccaa gctgatctgg gagtcagtga
     3781 gcgtcacagt ggtcgcagcc gtggaagcta tgaattggct gaagtccgct gcaaaactgc
     3841 tggccgctga ggtgaaggac aagaaaactg gcgaaattct gaggaaaaga tgcgccgtcc
     3901 actgggtgac ccctgatgga ttcccagtgt ggcaggagta taagaaaccc atccagacca
     3961 gactgaacct gatgttcctg ggccagtttc ggctgcagcc tacaatcaac actaataagg
     4021 acagtgagat tgatgctcat aaacaggaat cagggattgc acctaatttt gtgcacagcc
     4081 aggacggctc ccatctgcgg aagactgtgg tctgggctca cgagaaatac ggcatcgaat
     4141 ccttcgcact gattcatgac tcttttggaa ccatcccagc cgatgcagcc aacctgttca
     4201 aggctgtccg cgagactatg gtggacacct acgaaagttg tgatgtgctg gccgacttct
     4261 atgatcagtt tgctgaccag ctgcacgagt cacagctgga taagatgccc gcactgcctg
     4321 ccaaaggcaa cctgaatctg agagacatcc tggagtccga tttcgcattt gcctgactcg
     4381 aggatctttt tccctctgcc aaaaattatg gggacatcat gaagcccctt gagcatctga
     4441 cttctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat tttttgtgtc
     4501 tctcactcgg aaggacatat gggagggcaa atcatttaaa acatcagaat gagtatttgg
     4561 tttagagttt ggcaacatat gcccatatgc tggctgccat gaacaaaggt tggctataaa
     4621 gaggtcatca gtatatgaaa cagccccctg ctgtccattc cttattccat agaaaagcct
     4681 tgacttgagg ttagattttt tttatatttt gttttgtgtt atttttttct ttaacatccc
     4741 taaaattttc cttacatgtt ttactagcca gatttttcct cctctcctga ctactcccag
     4801 tcatagctgt ccctcttctc ttatggagat ccctcgacct gcagcccaag cttggcgtaa
     4861 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     4921 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     4981 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gcggatccgc
     5041 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     5101 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     5161 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     5221 taggcttttg caaaaagcta acttgtttat tgcagcttat aatggttaca aataaagcaa
     5281 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     5341 caaactcatc aatgtatctt atcatgtctg gatccgctgc attaatgaat cggccaacgc
     5401 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     5461 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     5521 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     5581 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     5641 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     5701 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5761 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     5821 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5881 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5941 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     6001 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     6061 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     6121 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     6181 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     6241 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     6301 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     6361 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     6421 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     6481 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     6541 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     6601 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     6661 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     6721 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     6781 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     6841 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     6901 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     6961 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     7021 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     7081 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     7141 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     7201 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     7261 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     7321 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgggtcgac attgattatt
     7381 g
//
