LOCUS       sequence-248073-        7586 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 1 vector. With lacZ as alternative cloning reporter.OriV 9 
            (pBBR322/ROP; medium copy number). Constitutive dCas9 in downstream 
            secondary site..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 91 to 125)
  AUTHORS   Valenzuela-Ortega M, French C
  TITLE     Joint universal modular plasmids (JUMP): a flexible vector platform 
            for synthetic biology.
  JOURNAL   Synth Biol (Oxf). 2021 Feb 2;6(1):ysab003. doi: 
            10.1093/synbio/ysab003. eCollection 2021.
  PUBMED    33623824
REFERENCE   2  (bases 1 to 7586)
  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"; journalName: "Synth Biol 
            (Oxf)."; date: "2021-02-2"; volume: "6(1):ysab003. doi"; pages: " 
            10.1093/synbio/ysab003. eCollection 2021"
COMMENT     Sequence Label: pJUMP29[dCas9]-1A(lacZ)
FEATURES             Location/Qualifiers
     source          1..7586
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    47..68
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     promoter        91..125
                     /label=﻿BBa_J23100
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 1.0
                     "
                     /citation=[1]
     RBS             130..150
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     misc_feature    486..506
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     promoter        511..545
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     RBS             549..569
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             join(586..609,610..612,613..3099,3100..3102,3103..4692)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGDS"
     terminator      4704..4731
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      4755..4849
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(4970..4989)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4981..5796
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     oriT            5963..6071
                     /note="incP origin of transfer"
     rep_origin      6107..6695
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6596..6615
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6849..6866
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6881..7021
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6947..6966)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7107..7129
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(7123..7314)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     terminator      complement(7494..7580)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ttaattaagg agttttgcag gtgccttgga acacctgctt ttcgctgaat tcgcggccgc
       61 ttctagagcg tctctggagt gagaccggag ttgacggcta gctcagtcct aggtacagtg
      121 ctagcgtact tgtttaactt taagaaggag atatacaatg gtagccggtg acccgaaact
      181 gctgctggaa gttccgcacg ctatctcttc ttctccgggt aactctctgg ctgttgttct
      241 gcaacgtcgt gactgggaaa acccgggtgt tacccagctg aaccgtctgg ctgctcaccc
      301 gccgttcgct tcttggcgta actctgaaga agctcgtacc gaccgtccgt cccagcagct
      361 gcgttctctg atccgtctgc tgaccaaacc ggaacgtaaa ctgtcttggc tgctgccgcc
      421 gctgtctaac aacggttcgt aacttctgac tgagttgcac gctggtctct cgctaatgtg
      481 agacgtacta gtagcggccg ctgcagggag tttacggcta gctcagtcct aggtatagtg
      541 ctagctactt gtttaacttt aagaaggaga tatacaatgg tagccgataa gaaatactca
      601 ataggcttag ctatcggcac aaatagcgtc ggatgggcgg tgatcactga tgaatataag
      661 gttccgtcta aaaagttcaa ggttctggga aatacagacc gccacagtat caaaaaaaat
      721 cttatagggg ctcttttatt tgacagtgga gagacagcgg aagcgactcg tcttaaacgg
      781 acagctcgta gaaggtatac acgtcggaag aatcgtattt gttatctaca ggagattttt
      841 tcaaatgaga tggcgaaagt agatgatagt ttctttcatc gacttgaaga gtcttttttg
      901 gtggaagaag ataagaagca tgaacgtcat cctatttttg gaaatatagt agatgaagtt
      961 gcttatcatg agaaatatcc aactatctat catctgcgaa aaaaattggt agattctact
     1021 gataaagcgg atttgcgctt aatctatttg gccttagcgc atatgattaa gtttcgtggt
     1081 cattttttga ttgagggaga tttaaatcct gataatagtg atgtggacaa actatttatc
     1141 cagttggtac aaacctacaa tcaattattt gaagaaaacc ctattaacgc aagtggagta
     1201 gatgctaaag cgattctttc tgcacgattg agtaaatcaa gacgattaga aaatctcatt
     1261 gctcagctcc ccggtgagaa gaaaaatggc ttatttggga atctcattgc tttgtcattg
     1321 ggtttgaccc ctaattttaa atcaaatttt gatttggcag aagatgctaa attacagctt
     1381 tcaaaagata cttacgatga tgatttagat aatttattgg cgcaaattgg agatcaatat
     1441 gctgatttgt ttttggcagc taagaattta tcagatgcta ttttactttc agatatccta
     1501 agagtaaata ctgaaataac taaggctccc ctatcagctt caatgattaa acgctacgat
     1561 gaacatcatc aagacttgac tcttttaaaa gctttagttc gacaacaact tccagaaaag
     1621 tataaagaaa tcttttttga tcaatcaaaa aacggatatg caggttatat tgatggggga
     1681 gctagccaag aagaatttta taaatttatc aaaccaattt tagaaaaaat ggatggtact
     1741 gaggaattat tggtgaaact aaatcgtgaa gatttgctgc gcaagcaacg gacctttgac
     1801 aacggctcta ttccccatca aattcacttg ggtgagctgc atgctatttt gagaaggcaa
     1861 gaagattttt atccattttt aaaagacaat cgtgagaaga ttgaaaaaat cttgactttt
     1921 cgcattcctt attatgttgg tccattggcg cgtggcaata gtcgttttgc atggatgact
     1981 cggaagtctg aagaaacaat taccccatgg aattttgaag aagttgtcga taaaggtgct
     2041 tcagctcaat catttattga acgcatgaca aactttgata aaaatcttcc aaatgaaaaa
     2101 gtactaccaa aacatagttt gctttatgag tattttacgg tttataacga attgacaaag
     2161 gtcaaatatg ttactgaagg aatgcgaaaa ccagcatttc tttcaggtga acagaagaaa
     2221 gccattgttg atttactctt caaaacaaat cgaaaagtaa ccgttaagca attaaaagaa
     2281 gattatttca aaaaaataga atgttttgat agtgttgaaa tttcaggagt tgaagataga
     2341 tttaatgctt cattaggtac ctaccatgat ttgctaaaaa ttattaaaga taaagatttt
     2401 ttggataatg aagaaaatga agatatctta gaggatattg ttttaacatt gaccttattt
     2461 gaagataggg agatgattga ggaaagactt aaaacatatg ctcacctctt tgatgataag
     2521 gtgatgaaac agcttaaacg tcgccgttat actggttggg gacgtttgtc tcgaaaattg
     2581 attaatggta ttagggataa gcaatctggc aaaacaatat tagatttttt gaaatcagat
     2641 ggttttgcca atcgcaattt tatgcagctg atccatgatg atagtttgac atttaaagaa
     2701 gatattcaaa aagcacaagt gtctggacaa ggcgatagtt tacatgaaca tattgcaaat
     2761 ttagctggta gccctgctat taaaaaaggt attttacaga ctgtaaaagt tgttgatgaa
     2821 ttggtcaaag taatggggcg gcataagcca gaaaatatcg ttattgaaat ggcacgtgaa
     2881 aatcagacaa ctcaaaaggg ccagaaaaat tcgcgagagc gtatgaaacg aatcgaagaa
     2941 ggtatcaaag aattaggaag tcagattctt aaagagcatc ctgttgaaaa tactcaattg
     3001 caaaatgaaa agctctatct ctattatctc caaaatggaa gagacatgta tgtggaccaa
     3061 gaattagata ttaatcgttt aagtgattat gatgtcgatg ccattgttcc acaaagtttc
     3121 cttaaagacg attcaataga caataaggtc ttaacgcgtt ctgataaaaa tcgtggtaaa
     3181 tcggataacg ttccaagtga agaagtagtc aaaaagatga aaaactattg gagacaactt
     3241 ctaaacgcca agttaatcac tcaacgtaag tttgataatt taacgaaagc tgaacgtgga
     3301 ggtttgagtg aacttgataa agctggtttt atcaaacgcc aattggttga aactcgccaa
     3361 atcactaagc atgtggcaca aattttggat agtcgcatga atactaaata cgatgaaaat
     3421 gataaactta ttcgagaggt taaagtgatt accttaaaat ctaaattagt ttctgacttc
     3481 cgaaaagatt tccaattcta taaagtacgt gagattaaca attaccatca tgcccatgat
     3541 gcgtatctaa atgccgtcgt tggaactgct ttgattaaga aatatccaaa acttgaatcg
     3601 gagtttgtct atggtgatta taaagtttat gatgttcgta aaatgattgc taagtctgag
     3661 caagaaatag gcaaagcaac cgcaaaatat ttcttttact ctaatatcat gaacttcttc
     3721 aaaacagaaa ttacacttgc aaatggagag attcgcaaac gccctctaat cgaaactaat
     3781 ggggaaactg gagaaattgt ctgggataaa gggcgagatt ttgccacagt gcgcaaagta
     3841 ttgtccatgc cccaagtcaa tattgtcaag aaaacagaag tacagacagg cggattctcc
     3901 aaggagtcaa ttttaccaaa aagaaattcg gacaagctta ttgctcgtaa aaaagactgg
     3961 gatccaaaaa aatatggtgg ttttgatagt ccaacggtag cttattcagt cctagtggtt
     4021 gctaaggtgg aaaaagggaa atcgaagaag ttaaaatccg ttaaagagtt actagggatc
     4081 acaattatgg aaagaagttc ctttgaaaaa aatccgattg actttttaga agctaaagga
     4141 tataaggaag ttaaaaaaga cttaatcatt aaactaccta aatatagtct ttttgagtta
     4201 gaaaacggtc gtaaacggat gctggctagt gccggagaat tacaaaaagg aaatgagctg
     4261 gctctgccaa gcaaatatgt gaatttttta tatttagcta gtcattatga aaagttgaag
     4321 ggtagtccag aagataacga acaaaaacaa ttgtttgtgg agcagcataa gcattattta
     4381 gatgagatta ttgagcaaat cagtgaattt tctaagcgtg ttattttagc agatgccaat
     4441 ttagataaag ttcttagtgc atataacaaa catagagaca aaccaatacg tgaacaagca
     4501 gaaaatatta ttcatttatt tacgttgacg aatcttggag ctcccgctgc ttttaaatat
     4561 tttgatacaa caattgatcg taaacgatat acgtctacaa aagaagtttt agatgccact
     4621 cttatccatc aatccatcac tggtctttat gaaacacgca ttgatttgag tcagctagga
     4681 ggtgattcgt gatcgatcac actggctcac cttcgggtgg gcctttctgc gtttatatgt
     4741 tcgctctagt cttggactcc tgttgataga tccagtaatg acctcagaac tccatctgga
     4801 tttgttcaga acgctcggtt gccgccgggc gttttttatt ggtgagaatc caggggtccc
     4861 caataattac gatttaaatt tgtgtctcaa aatctctgat gttacattgc acaagataaa
     4921 aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt
     4981 atgagccata ttcagcgtga aacgagctgt agccgtccgc gtctgaacag caacatggat
     5041 gcggatctgt atggctataa atgggcgcgt gataacgtgg gtcagagcgg cgcgaccatt
     5101 tatcgtctgt atggcaaacc ggatgcgccg gaactgtttc tgaaacatgg caaaggcagc
     5161 gtggcgaacg atgtgaccga tgaaatggtg cgtctgaact ggctgaccga atttatgccg
     5221 ctgccgacca ttaaacattt tattcgcacc ccggatgatg cgtggctgct gaccaccgcg
     5281 attccgggca aaaccgcgtt tcaggtgctg gaagaatatc cggatagcgg cgaaaacatt
     5341 gtggatgcgc tggccgtgtt tctgcgtcgt ctgcatagca ttccggtgtg caactgcccg
     5401 tttaacagcg atcgtgtgtt tcgtctggcc caggcgcaga gccgtatgaa caacggcctg
     5461 gtggatgcga gcgattttga tgatgaacgt aacggctggc cggtggaaca ggtgtggaaa
     5521 gaaatgcata aactgctgcc gtttagcccg gatagcgtgg tgacccacgg cgattttagc
     5581 ctggataacc tgattttcga tgaaggcaaa ctgattggct gcattgatgt gggccgtgtg
     5641 ggcattgcgg atcgttatca ggatctggcc attctgtgga actgcctggg cgaatttagc
     5701 ccgagcctgc aaaaacgtct gtttcagaaa tatggcattg ataatccgga tatgaacaaa
     5761 ctgcaatttc atctgatgct ggatgaattt ttctaataat taattggacc gcggtccgcg
     5821 cgttgtcctt ttccgctgca taaccctgct tcggggtcat tatagcgatt ttttcggtat
     5881 atccatcctt tttcgcacga tatacaggat tttgccaaag ggttcgtgta gactttcctt
     5941 ggtgtatcca acggcgtcag ccgggcagga taggtgaagt aggcccaccc gcgagcgggt
     6001 gttccttctt cactgtccct tattcgcacc tggcggtgct caacgggaat cctgctctgc
     6061 gaggctggcc gtaggccggc cccgtagaaa agatcaaagg atcttcttga gatccttttt
     6121 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     6181 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     6241 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     6301 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     6361 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     6421 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     6481 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     6541 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     6601 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     6661 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     6721 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     6781 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     6841 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc
     6901 ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg
     6961 atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc
     7021 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     7081 cgcttacaga caagctgtga cagtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     7141 atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gcagcgattc
     7201 acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt
     7261 ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcactgatgc
     7321 ctccgtgtaa gggggatttc tgttcatggg ggtaatgata ccgatgaaac gagagaggat
     7381 gctcacgata cgggttactg atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa
     7441 acaactggcg gtatggatgc ggcgggggcg cgcccagctg tctagggcgg cggatttgtc
     7501 ctactcagga gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtctttcgac
     7561 tgagcctttc gttttatttg atgcct
//
