LOCUS       sequence-244343-        7276 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ABA-inducible split T7 gRNA for GFP vector using RNAPN N-29-1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7276)
  AUTHORS   Pu J, Kentala K, Dickinson BC
  TITLE     Multidimensional Control of Cas9 by Evolved RNA Polymerase-Based 
            Biosensors.
  JOURNAL   ACS Chem Biol. 2018 Feb 16;13(2):431-437. doi: 
            10.1021/acschembio.7b00532. Epub 2017 Aug 22.
  PUBMED    28809467
REFERENCE   2  (bases 1 to 7276)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1021/acschembio.7b00532"; journalName: "ACS Chem Biol"; date: 
            "2018-02-16- 16"; volume: "13"; issue: "2"; pages: "431-437"
COMMENT     Sequence Label: pJin-239
FEATURES             Location/Qualifiers
     source          1..7276
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    465..748
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     primer_bind     complement(944..963)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(945..963)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        989..1088
                     /label=CMVd1 promoter
                     /note="truncated human cytomegalovirus (CMV) immediate 
                     early promoter that yields slightly reduced expression 
                     relative to the full-length promoter"
     promoter        3862..3961
                     /label=CMVd1 promoter
                     /note="truncated human cytomegalovirus (CMV) immediate 
                     early promoter that yields slightly reduced expression 
                     relative to the full-length promoter"
     primer_bind     complement(5496..5514)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    5551..5672
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5562..5581)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5616..5635
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             6071..6865
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MLEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6125..6144)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6735..6754
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(join(7037..7276,1..349))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7117..7136)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
       61 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
      121 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
      181 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
      241 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
      301 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatttcaag aagatccttt
      361 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
      421 catgagatta tcaaaaagga tcttcaccta gatcctttta tagtccggaa atacaggaac
      481 gcacgctgga tggcccttcg ctgggatggt gaaaccatga aaaatggcag cttcagtgga
      541 ttaagtgggg gtaatgtggc ctgtaccctc tggttgcata ggtattcata cggttaaaat
      601 ttatcaggcg cgattgcggc agtttttcgg gtggtttgtt gccattttta cctgtctgct
      661 gccgtgatcg cgctgaacgc gttttagcgg tgcgtacaat taagggatta tggtaaatcc
      721 acttactgtc tgccctcgta gccatcgaga taaaccgcag tactccggcc acgatggaaa
      781 aaaactaaca gatactctag tagaaaaaaa ctaacagata ctctagtaaa aaaaatctcg
      841 ccaacaagtt gacgagataa acacggcatt ttgccttgtt ttagtagatt ctgtaatttt
      901 cattacagag tactaaaacg ccgaggtgaa gttcgagggc tctccctata gtgagtcgta
      961 ttacgtccgg cgtagaggat cgagatctcc aaaatcaacg ggactttcca aaatgtcgta
     1021 ataaccccgc cccgttgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     1081 gcatagctgg tttagtgaac cgtcagatca ctagaagctt acgctaccac catgacccag
     1141 gacgagttta cccagctgtc acagtcaatc gccgagtttc acacctatca gctggggaac
     1201 ggaagatgta gtagtctgct ggcccagaga atccacgctc cccctgaaac cgtgtggagc
     1261 gtggtccgga gattcgatag accacaaatc tacaagcact tcatcaagag ctgcaacgtg
     1321 tccgaagatt tcgagatgcg cgtcggatgt acacgagacg tgaacgtcat ctctggcctg
     1381 cccgcaaata ctagtcgaga gcggctggac ctgctggacg atgacaggcg cgtgactggg
     1441 ttctccatta ccggcgggga acacagactg aggaactaca agtctgtgac cacagtccat
     1501 aggtttgaga aagaggaaga ggaagagcgc atttggaccg tggtcctgga aagctacgtg
     1561 gtcgatgtgc ccgagggcaa tagcgaagag gacacaaggc tgtttgccga tactgtgatt
     1621 cggctgaacc tgcagaaact ggcaagcatt actgaggcta tgaacacgtc gggtggaagc
     1681 ggtaaagcat ttatgcaagt tgtcgaggct gacatgctct ctaagggtct actcggtggc
     1741 gaggcgtggt cttcgtggca taaggaagac tctattcatg taggagtacg ctgcatcgag
     1801 atgctcattg agtcaaccgg aatggttagc ttacaccgcc aaaatgctgg cgtagtaggt
     1861 caagactctg agactatcga actcgcacct gaatacgctg aggctatcgc aacccgtgca
     1921 ggtgcgctgg ctggcatctc tccgatgttc caaccttgcg tagttcctcc taagccgtgg
     1981 actggcatta ctggtggtgg ctattgggct aacggtcgtc gtcctctggc gctggtgcgt
     2041 actcacagta agaaagcact gatgcgctac gaagacgttt acatgcctga ggtgtacaaa
     2101 gcgattaaca ttgcgcaaaa caccgcatgg aaaatcaaca agaaagtcct agcggtcgcc
     2161 aacgtaatca ccaagtggaa gcattgtccg gtcgaggaca tccctgcgat tgagcgtgaa
     2221 gaactcccga tgaaaccgga agacatcgac atgaatcctg aggctctcac cgcgtggaaa
     2281 cgtgctgccg ctgctgtgta ccgcaaggac aaggctcgca agtctcgccg tatcagcctt
     2341 gagttcatgc ttgagcaagc caataagttt gctaaccata aggccatctg gttcccttac
     2401 aacatggact ggcgcggtcg tgtttacgct gtgtcaatgt tcaacccgca aggtaacgat
     2461 atgaccaaag gactgcttac gctggcgaaa ggtaaaccaa tcggtaagga aggttactac
     2521 tggctgaaaa tccacggtgc aaactgtgcg ggtgtcgata aggttccgtt ccctgagcgc
     2581 atcaagttca ttgaggaaaa ccacgagaac atcatggctt gcgctaagtc tccactggag
     2641 aacacttggt gggctgagca agattctccg ttctgcttcc ttgcgttctg ctttgagtac
     2701 gctggggtac agcaccacgg cctgagctat aactgctccc ttccgctggc gtttgacggg
     2761 tcttgctctg gcatccagca cttctccgcg atgctccgag atgaggtagg tggtcgcgcg
     2821 gttaacttgc ttcctagtga aaccgttcag gacatctacg ggattgttgc taagaaagtc
     2881 aacgagattc tacaagcaga cgcaatcaat gggaccgata acgaagtagt taccgtgacc
     2941 gatgagaaca ctggtgaaat ctctgagaaa gtcaagctgg gcactaaggc actggctggt
     3001 caatggctgg cttacggtgt tactcgcagt gtgactaagc gttcagtcat gacgctggct
     3061 tacgggtcca aagagttcgg cttccgtcaa caagtgctgg aagataccat tcagccagct
     3121 attgattccg gcaagggtct gatgttcact cagccgaatc aggctgctgg atacatggct
     3181 aagctgattt gggaatctgt gagcgtgacg gtggtagctg cggttgaagc aatgaactgg
     3241 cttaagtctg ctgctaagct gctggctgct gaggtcaaag ataagaagac tggagagatt
     3301 cttcgcaagc gttgcgctgt gcattgggta actcctgatg gtttccctgt gtggcaggaa
     3361 tacaagaagc ctattcagac gcgcttgaac ctgatgttcc tcggtcagtt ccgcttacag
     3421 cctaccatta acaccaacaa agatagcgag attgatgcac acaaacagga gtctggtatc
     3481 gctcctaact ttgtacacag ccaagacggt agccaccttc gtaagactgt agtgtgggca
     3541 cacgagaagt acggaatcga atcttttgca ctgattcacg actccttcgg taccattccg
     3601 gctgacgctg cgaacctgtt caaagcagtg cgcgaaacta tggttgacac atatgagtct
     3661 tgtgatgtac tggctgattt ctacgaccag ttcgctgacc agttgcacga gtctcaattg
     3721 gacaaaatgc cagcacttcc ggctaaaggt aacttgaacc tccgtgacat cttagagtcg
     3781 gacttcgcgt tcgcgtaatg tttaaacgaa ttcgggctcg gtacccgggg atcctctaga
     3841 gtcgacctgc aggcatgcaa gccaaaatca acgggacttt ccaaaatgtc gtaataaccc
     3901 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
     3961 tggtttagtg aaccgtcaga tcactagaag cttacgctac caccatgaac acgattaaca
     4021 tcgctaagaa cgacttctct gacatcgaac tggctgctat cccgttcaac actctggctg
     4081 accattacgg tgagcgttca gctcgcggac agttggccct tgagcatgag tcttacgaga
     4141 tgggtgaagc acgcttccgc aagatgtttg agcgtcaact taaagctggt gaggttgcgg
     4201 ataacgctgc cgccaagcct ctcatcacta ccctactccc taagatgatt gcccgcatca
     4261 acgactggtt tgaggaagtg aaagctaagc gcggcaggcg cccgacagcc ttccagttcc
     4321 taaaagaaat caagccggaa gccgtagcgt acatcaccat taagacctct ctggcttgcc
     4381 taaccagtgc tgacaataca accgttcagg ctgtagcaag cgcaatcggt cggaccattg
     4441 aggacgaggc tcgcttcggt cgtatccgtg accttgaagc taagcacttc aagaaaaacg
     4501 ttgaggaaca actcaacaag cgcgtagggc acgtctacaa gggtggctct ggctctggct
     4561 cgagcgtccc cctgtatggg ttcaccagca tttgtgggcg gagaccagag atggaggcag
     4621 ccgtcagcac tattccccgg ttcctgcagt caagctccgg cagcatgctg gacgggaggt
     4681 tcgatccaca gtccgccgct cacttctttg gggtctacga tggacatggc gggtcccagg
     4741 tggccaacta ttgcagggag cgcatgcacc tggctctggc agaggaaatc gccaaggaga
     4801 aacctatgct gtgcgacgga gatacatggc tggaaaagtg gaagaaagcc ctgttcaact
     4861 cttttctgcg ggtggactcc gagatcgaat ctgtcgctcc tgagaccgtg ggctctacaa
     4921 gtgtggtcgc agtggtcttc ccatctcaca tttttgtcgc aaattgcggc gacagccggg
     4981 ccgtgctgtg caggggaaag accgccctgc cactgagcgt ggaccataaa cccgacaggg
     5041 aggatgaagc agcccgcatc gaggctgcag gaggcaaagt gatccagtgg aacggagcac
     5101 gggtgtttgg cgtcctggcc atgtcacgca gcattgggga tcgatacctg aaaccatcaa
     5161 tcattcccga ccctgaagtg actgccgtca agagagtgaa agaagacgat tgcctgatcc
     5221 tggctagcga cggcgtctgg gatgtgatga ccgacgagga agcatgtgag atggcccgaa
     5281 agcggattct gctgtggcat aagaaaaatg ccgtggctgg cgatgctagt ctgctggcag
     5341 acgagcggag aaaggaaggg aaagatcccg ccgctatgag tgcagccgaa tatctgtcaa
     5401 aactggctat tcagaggggc agtaaggaca acatctccgt cgtcgtcgtg gacctgaagt
     5461 gaaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc
     5521 ccttggggcg gccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca
     5581 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     5641 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     5701 ttcaggttca gggggagatg tgggaggttt ttttaagcaa gtaaaacctc tacaaatgtg
     5761 gtaaaatcga attctaatgg atcctctttg cgcttgcgtt ttcccttgtc cagatagccc
     5821 agtagctgac attcatccgg ggtcagcacc atttctgcgg actggctttc tacgtgttcc
     5881 gcttccttta gcagcccttg cgccctgagt gcttgcggca gcgtgagctt caaaagaatt
     5941 gccagctggg gcgccctctg gtaaggttgg gaagccctgc aaagtaaact ggatggcttt
     6001 cttgccgcca aggatctgat ggcgcagggg atcaagatct gatcaagaga caggatgacg
     6061 gtcgtttcgc atgcttgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     6121 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     6181 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     6241 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     6301 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     6361 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     6421 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     6481 gaaacatcgc atcgagcgag cacgcactcg gatggaagcc ggtcttgtcg atcaggatga
     6541 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     6601 tatgccggat ggtgaggatc tcgtcgtgac tcatggcgat gcctgcttgc cgaatatcat
     6661 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     6721 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     6781 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     6841 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     6901 acgcccaacc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     6961 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     7021 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     7081 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     7141 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     7201 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     7261 tgtaggtcgt tcgctc
//
