p6XHis-T7(P266L)

LOCUS       p6XHis-T7(P266L)                5428 bp ds-DNA     circular SYN 03-NOV-2021
DEFINITION  Expresses a T7 RNA Polymerase that produces long RNA transcripts in 
            high yield and generates fewer abortive transcripts..
ACCESSION   .
VERSION     .
KEYWORDS    p6XHis-T7(P266L)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5428)
  AUTHORS   Chillon I, Marcia M, Legiewicz M, Liu F, Somarowthu S, Pyle AM
  TITLE     Native Purification and Analysis of Long RNAs.
  JOURNAL   Methods Enzymol. 2015;558:3-37. doi: 10.1016/bs.mie.2015.01.008. 
            Epub 2015 Feb 27.
  PUBMED    26068736
REFERENCE   2  (bases 1 to 5428)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 3, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5428
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             13..30
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             40..57
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     promoter        complement(3084..3112)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(3139..3157)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3225..3329
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3330..4190
                     /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     complement(3548..3567)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4361..4949
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4850..4869
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5103..5120
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5237..5258
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5273..5303
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5311..5327
                     /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)."
     primer_bind     5316..5338
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5335..5351
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5335..5351
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     RBS             5400..5422
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat
       61 atgctcgaga tgaacacgat taacatcgct aagaacgact tctctgacat cgaactggct
      121 gctatcccgt tcaacactct ggctgaccat tacggtgagc gtttagctcg cgaacagttg
      181 gcccttgagc atgagtctta cgagatgggt gaagcacgct tccgcaagat gtttgagcgt
      241 caacttaaag ctggtgaggt tgcggataac gctgccgcca agcctctcat cactacccta
      301 ctccctaaga tgattgcacg catcaacgac tggtttgagg aagtgaaagc taagcgcggc
      361 aagcgcccga cagccttcca gttcctgcaa gaaatcaagc cggaagccgt agcgtacatc
      421 accattaaga ccactctggc ttgcctaacc agtgctgaca atacaaccgt tcaggctgta
      481 gcaagcgcaa tcggtcgggc cattgaggac gaggctcgct tcggtcgtat ccgtgacctt
      541 gaagctaagc acttcaagaa aaacgttgag gaacaactca acaagcgcgt agggcacgtc
      601 tacaagaaag catttatgca agttgtcgag gctgacatgc tctctaaggg tctactcggt
      661 ggcgaggcgt ggtcttcgtg gcataaggaa gactctattc atgtaggagt acgctgcatc
      721 gagatgctca ttgagtcaac cggaatggtt agcttacacc gccaaaatgc tggcgtagta
      781 ggtcaagact ctgagactat cgaactcgca cctgaatacg ctgaggctat cgcaacccgt
      841 gcaggtgcgc tggctggcat ctctctgatg ttccaacctt gcgtagttcc tcctaagccg
      901 tggactggca ttactggtgg tggctattgg gctaacggtc gtcgtcctct ggcgctggtg
      961 cgtactcaca gtaagaaagc actgatgcgc tacgaagacg tttacatgcc tgaggtgtac
     1021 aaagcgatta acattgcgca aaacaccgca tggaaaatca acaagaaagt cctagcggtc
     1081 gccaacgtaa tcaccaagtg gaagcattgt ccggtcgagg acatccctgc gattgagcgt
     1141 gaagaactcc cgatgaaacc ggaagacatc gacatgaatc ctgaggctct caccgcgtgg
     1201 aaacgtgctg ccgctgctgt gtaccgcaag gacaaggctc gcaagtctcg ccgtatcagc
     1261 cttgagttca tgcttgagca agccaataag tttgctaacc ataaggccat ctggttccct
     1321 tacaacatgg actggcgcgg tcgtgtttac gctgtgtcaa tgttcaaccc gcaaggtaac
     1381 gatatgacca aaggactgct tacgctggcg aaaggtaaac caatcggtaa ggaaggttac
     1441 tactggctga aaatccacgg tgcaaactgt gcgggtgtcg ataaggttcc gttccctgag
     1501 cgcatcaagt tcattgagga aaaccacgag aacatcatgg cttgcgctaa gtctccactg
     1561 gagaacactt ggtgggctga gcaagattct ccgttctgct tccttgcgtt ctgctttgag
     1621 tacgctgggg tacagcacca cggcctgagc tataactgct cccttccgct ggcgtttgac
     1681 gggtcttgct ctggcatcca gcacttctcc gcgatgctcc gagatgaggt aggtggtcgc
     1741 gcggttaact tgcttcctag tgaaaccgtt caggacatct acgggattgt tgctaagaaa
     1801 gtcaacgaga ttctacaagc agacgcaatc aatgggaccg ataacgaagt agttaccgtg
     1861 accgatgaga acactggtga aatctctgag aaagtcaagc tgggcactaa ggcactggct
     1921 ggtcaatggc tggcttacgg tgttactcgc agtgtgacta agcgttcagt catgacgctg
     1981 gcttacgggt ccaaagagtt cggcttccgt caacaagtgc tggaagatac cattcagcca
     2041 gctattgatt ccggcaaggg tctgatgttc actcagccga atcaggctgc tggatacatg
     2101 gctaagctga tttgggaatc tgtgagcgtg acggtggtag ctgcggttga agcaatgaac
     2161 tggcttaagt ctgctgctaa gctgctggct gctgaggtca aagataagaa gactggagag
     2221 attcttcgca agcgttgcgc tgtgcattgg gtaactcctg atggtttccc tgtgtggcag
     2281 gaatacaaga agcctattca gacgcgcttg aacctgatgt tcctcggtca gttccgctta
     2341 cagcctacca ttaacaccaa caaagatagc gagattgatg cacacaaaca ggagtctggt
     2401 atcgctccta actttgtaca cagccaagac ggtagccacc ttcgtaagac tgtagtgtgg
     2461 gcacacgaga agtacggaat cgaatctttt gcactgattc acgactcctt cggtaccatt
     2521 ccggctgacg ctgcgaacct gttcaaagca gtgcgcgaaa ctatggttga cacatatgag
     2581 tcttgtgatg tactggctga tttctacgac cagttcgctg accagttgca cgagtctcaa
     2641 ttggacaaaa tgccagcact tccggctaaa ggtaacttga acctccgtga catcttagag
     2701 tcggacttcg cgttcgcgta acgccaaatc aatacgactc cggatccaca ggacgggtgt
     2761 ggtcgccatg atcgcgtagt cgatagtggc tccaagtagc gaagcgagca ggactgggcg
     2821 gcggccaaag cggtcggaca gtgctccgag aacgggtgcg catagaaatt gcatcaacgc
     2881 atatagcgct agcagcacgc catagtgact ggcgatgctg tcggaatgga cgatatcccg
     2941 caagaggccc ggcagtaccg gcataaccaa gcctatgcct acagcatcca gggtgacggt
     3001 gccgaggatg acgatgagcg cattgttaga tttcatacac ggtgcctgac tgcgttagca
     3061 atttaactgt gataaactac cgcattaaag cttatcgatg ataagctgtc aaacatgaga
     3121 attcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     3181 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     3241 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     3301 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     3361 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     3421 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     3481 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     3541 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     3601 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     3661 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     3721 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     3781 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     3841 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     3901 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     3961 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     4021 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     4081 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     4141 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     4201 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     4261 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     4321 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgt
     4381 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     4441 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     4501 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     4561 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     4621 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     4681 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     4741 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     4801 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     4861 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     4921 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     4981 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     5041 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     5101 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     5161 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     5221 gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact
     5281 ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
     5341 acagctatga ccatgattac gccaagcttg catgcctgca ggtcgactct agaaataatt
     5401 ttgtttaact ttaagaagga gatatacc
//