LOCUS       sequence-227927-        8952 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains intB13 integrase casette for knock-in to pCAP-BAC backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8952)
  AUTHORS   Bauman KD, Li J, Murata K, Mantovani SM, Dahesh S, Nizet V, Luhavaya
            H, Moore BS
  TITLE     Refactoring the Cryptic Streptophenazine Biosynthetic Gene Cluster 
            Unites Phenazine, Polyketide, and Nonribosomal Peptide Biochemistry.
  JOURNAL   Cell Chem Biol. 2019 Feb 15. pii: S2451-9456(19)30038-8. doi: 
            10.1016/j.chembiol.2019.02.004.
  PUBMED    30853419
REFERENCE   2  (bases 1 to 8952)
  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: "Cell Chem 
            Biol. 2019 Feb 15. pii: S2451-9456(19)30038-8. doi: 
            10.1016/j.chembiol.2019.02.004."
COMMENT     Sequence Label: pJZ001
FEATURES             Location/Qualifiers
     source          1..8952
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    3..27
                     /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)."
     CDS             83..100
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    1022..1038
                     /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)."
     CDS             complement(3510..4043)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(4232..4237,4238..4254,4255..4260))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     oriT            4274..4382
                     /note="incP origin of transfer"
     protein_bind    4522..4569
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             5310..5354
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(5392..5410)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5406..5453
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(5673..6332)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     6247..6266
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(6333..6435)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(6827..6844)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6961..7505)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(7041..7060)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7730..8812)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     8774..8793
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(8813..8890)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     join(8936..8952,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(8936..8952,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atgggcagca gccatcacca tcatcaccac agccaggatc cgaattcgag
      121 ctcggcgcgc ctgcaggtcg acaagctttc tatcgccttc ttgacgagtt cttctgaggt
      181 taactgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
      241 acctgggtcc ttttcatcac gtgctataaa aataattata atttaaattt tttaatataa
      301 atatataaat taaaaataga aagtaaaaaa agaaattaaa gaaaaaatag tttttgtttt
      361 ccgaagatgt aaaagactct agggggatcg ccaacaaata ctacctttta tcttgctctt
      421 cctgctctca ggtattaatg ccgaattgtt tcatcttgtc tgtgtagaag accacacacg
      481 aaaatcctgt gattttacat tttacttatc gttaatcgaa tgtatatcta tttaatctgc
      541 ttttcttgtc taataaatat atatgtaaag tacgcttttt gttgaaattt tttaaacctt
      601 tgtttatttt tttttcttca ttccgtaact cttctacctt ctttatttac tttctaaaat
      661 ccaaatacaa aacataaaaa taaataaaca cagagtaaat tcccaaatta ttccatcatt
      721 aaaagatacg aggcgcgtgt aagttacagg caagcggccg aattctgctg ggtgtgggtc
      781 tatggatgcc agtggaactc tttggacgat ctttgttggc ggggacgaaa aaataggcta
      841 caatctaggt cttcgctggc cacctcggcg aagaaagagc ttgggaatca acgacttact
      901 gccctagtaa acgtctcgtt tcccgctcca agttatgatc tacagacttc tactgacgtc
      961 tgtaagtcgt tgaaaagagg ggcttcggcc ccttttttca ttccagcgag cgccacggaa
     1021 attgtgagcg gataacaatt tccacccaca gccagccttt tttaggcgcc aatctaggca
     1081 caggaatacg ggtgaggcgg cttccggata gttgctgggg ctgagcggga catatgacga
     1141 gcatagggcc taagtcatta cttaggagcc tcgaaatggc actctctgat ctgatcgtcc
     1201 ggcaggccaa gaccaccggc aaacgctaca ccctctacga caacgactgc ctgagcctga
     1261 tggtctctgc cgcaggcggc aagtcttgga tgttccgcta ctcctggctg ggcaagcaaa
     1321 agcgcatggc cctgggcggc tatcctgccc tcagcctgcg cgaggcccgc gccgagcggg
     1381 acaaggccca ggccctgatc gccaggggga tcgatcccca aatcgaacgc gatcagcgtc
     1441 ggcacgcggc caagctggcc ggcgaataca ccttcaagaa cgtcttcgat gcctgggtcg
     1501 agcatcgccg caaggaactc aaggaaggcc gccagagtac gctttcgcag atcctgcgca
     1561 tcttcaacaa ggacgtgctg cccaccctag ggaagatgtc catctatgac attcgccggc
     1621 cccagctcct gggcgtcctg gcggcgatcg agaagcgcaa ggcgttcacc acggccgaga
     1681 aggtccgcac ctggttcaac cagatgttcc gctatgcctt ggtcatcgcc gaggggctgg
     1741 aagtcaaccc ggctgcggac ctggacgtgg tggccgagcc caagcccccg gtggcccaca
     1801 acccctatct gcacctgccc gaactgcccg agttccttca gaagctgcga cgctacaacc
     1861 cccggggctg gcagacccag cttggcgtgc ggctactgtt cctgacgggg gtgcgcacgg
     1921 gcgagctgcg gttggccgaa cccgagcagt tcgacctcga ccggggcttt tggatcatcc
     1981 cgccggaggt cgtcaagcaa ctccaggacg aaatgcgcaa ggccggcaag cggccgcagg
     2041 acgtgccgcc ctacatcgtg cccctgtccc tgcaggccat cgagatcgtg cgctatctcc
     2101 tgggcgtgat gcggccggcg cagaagtacc tgctgtcgca ccgcagcgaa ctcaagaaac
     2161 gcatcagcga gaacaccctc aacaaggccg tgcagctcat gggctacgag gggcgcctga
     2221 ccgggcacgg catccgcggc accatctcga cggcactcaa cgagatcggc tacccgaaga
     2281 tttgggtgga cgcgcagctt tcgcactccg accccaacaa ggtgagttcg gcctacaacc
     2341 acgccaagta cgtggagccg cgtcgccgca tgatgcagga ctgggccgat cgcctggacc
     2401 tgctcgaaca gggcgaagtg caagctgcga gcgcgcacct gaccatccgt atcgacgggg
     2461 tgccagcgat ggcggaagtg gaggaagcgg tggacgtggc cccgacggtc gccgagcccg
     2521 gtgtctccgg ctcaccgccc gtggcggcca cgcccatcgt cgtgaccccc aacagcggcg
     2581 gaatcacgtt ccagcgcctg tcgcaggtgc cgccgcctcc ggcgcatgct cctgagtcgg
     2641 aggtctctgc catccagcgc gagcgtgagg aaatgctggc catgtacgag tcgccgaaca
     2701 atctgccggt cgcgctgttc gggaagctgg ctgggaagtc caaagaccag atcaaccgcg
     2761 agctgaaggc aggcaagcta ctgtccatca gcttgggcaa ccgggggcag cgcgttcccg
     2821 actggcaact ggtgccgctc aagcgccggc tggcccaagc gctcatgaac caatgcccgc
     2881 atgtagattc ttgggcgctg tatcgtctgt tgaccaagcc ccattccaac ctcggaaacc
     2941 gtgcagccat cgatgtcgtc acaccgacca acgtgggcaa agtattgcaa gccgtcacgc
     3001 cctataagga gttcgagcgc tccagtacag acgagacacc ccagttctcc gagttcgttc
     3061 gccagttgca acgccacgtg aatgcgctcg aagaagcgcc gtgctgacga gccaggactt
     3121 cgacgccaac ggcgtgccgc aacgcgtgtt tacctagacg ctaggagtgc gttaaatccc
     3181 gtcgtgggcg gctattgcaa cgaactgatc aaccagacgg ccaagaccga ctgcaattcc
     3241 tcccggtaag gtttttcgcc tccgcgtccg gatgtccgtt ccttgagcgg cgggtaggag
     3301 gcctgtaccc aggatttatg aggttccccg ccccaaggtc cattcccatg tgcgaaccga
     3361 cagctcggca aacagacctg cctgtacgta ggggtcgtgg gccacgaagg cgtgcacgtc
     3421 gtcgatatgc gccgcctgca caagcggcgt tgtgacaatt taccgaacaa ctccgcggcc
     3481 gggaagccga tctcggcttg aacgaattgt taggtggcgg tacttgggtc gatatcaaag
     3541 tgcatcactt cttcccgtat gcccaacttt gtatagagag ccactgcggg atcgtcaccg
     3601 taatctgctt gcacgtagat cacataagca ccaagcgcgt tggcctcatg cttgaggaga
     3661 ttgatgagcg cggtggcaat gccctgcctc cggtgctcgc cggagactgc gagatcatag
     3721 atatagatct cactacgcgg ctgctcaaac ttgggcagaa cgtaagccgc gagagcgcca
     3781 acaaccgctt cttggtcgaa ggcagcaagc gcgatgaatg tcttactacg gagcaagttc
     3841 ccgaggtaat cggagtccgg ctgatgttgg gagtaggtgg ctacgtctcc gaactcacga
     3901 ccgaaaagat caagagcagc ccgcatggat ttgacttggt cagggccgag cctacatgtg
     3961 cgaatgatgc ccatacttga gccacctaac tttgttttag ggcgactgcc ctgctgcgta
     4021 acatcgttgc tgctgcgtaa catcgttgct gctccataac atcaaacatc gacccacggc
     4081 gtaacgcgct tgctgcttgg atgcccgagg catagactgt acaaaaaaac agtcataaca
     4141 agccatgaaa accgccactg cgccgttacc accgctgcgt tcggtcaagg ttctggacca
     4201 gttgcgtgag cgcatacgct acttgcatta cagtttacga accgaacagg cttatgtcaa
     4261 ttcggccggc ctacggccag cctcgcagag caggattccc gttgagcacc gccaggtgcg
     4321 aataagggac agtgaagaag gaacacccgc tcgcgggtgg gcctacttca cctatcctgc
     4381 ccggctgacg ccgttggata caccaaggaa agtctacacg aaccctttgg caaaatcctg
     4441 tatatcgtgc gaaaaaggat ggatataccg aaaaaatcgc tataatgacc ccgaagcagg
     4501 gttatgcagc ggaaaagaaa agaagttcct attccgaagt tcctattctc tagaaagtat
     4561 aggaacttca tgcatgatct gaagatcagc agttcaacct gttgatagta cgtactaagc
     4621 tctcatgttt cacgtactaa gctctcatgt ttaacgtact aagctctcat gtttaacgaa
     4681 ctaaaccctc atggctaacg ctccatcgta tccgtattcc tcttattttt tctctttctc
     4741 tatatccatt tctttctctc taggtgtgtc ctctctctct cttcaatttc tctactccgc
     4801 attccaacgc atccttcccc caacctccca tttcctcctt acggcccgat agcgatcgtc
     4861 tttccctcgc tatcactcgc taccggcccc tcctctgcac cgtaacctcc tacgtattta
     4921 ccatatcata aagttttttc cgacgcttat cgctgacccc ctgtcgccct cctattggct
     4981 tccggattat cttcttgtcc ataaggtgat ccatgcttcc tgaagattcc cgaaatgtgt
     5041 ccactttggc ggggaatcat tccatccact tctttctctc tcgctttcct cattcggcgc
     5101 tccccttccg cgtctcattg gtcttccgct ccgtttttgc tttgccgatg ttacttgggg
     5161 agaggtgcga taatcctttc gcaaaaactc ggtttgacgc ctcccatggt ataaatagtg
     5221 ggcgtttggt acggcggctc cgccacggca tcaatcggcc caatcggcaa atttgcggtc
     5281 aatcgtcttg cgcgaaactc gagtctggta aagaaaccgc tgctgcgaaa tttgaacgcc
     5341 agcacatgga ctcgtctact agcgcagctt aattaaccta ggctgctgcc accgctgagc
     5401 aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaac
     5461 ctcaggcatt tgagaagcac acggtcacac tgcttccggt agtcaataaa ccggtaaacc
     5521 agcaatagac ataagcggct atttaacgac cctgccctga accgacgacc gggtcgaatt
     5581 tgctttcgaa tttctgccat tcatccgctt attatcactt attcaggcgt agcaccaggc
     5641 gtttaagggc accaataact gccttaaaaa aattacgccc cgccctgcca ctcatcgcag
     5701 tactgttgta attcattaag cattctgccg acatggaagc catcacagac ggcatgatga
     5761 acctgaatcg ccagcggcat cagcaccttg tcgccttgcg tataatattt gcccatagtg
     5821 aaaacggggg cgaagaagtt gtccatattg gccacgttta aatcaaaact ggtgaaactc
     5881 acccagggat tggctgagac gaaaaacata ttctcaataa accctttagg gaaataggcc
     5941 aggttttcac cgtaacacgc cacatcttgc gaatatatgt gtagaaactg ccggaaatcg
     6001 tcgtggtatt cactccagag cgatgaaaac gtttcagttt gctcatggaa aacggtgtaa
     6061 caagggtgaa cactatccca tatcaccagc tcaccgtctt tcattgccat acggaactcc
     6121 ggatgagcat tcatcaggcg ggcaagaatg tgaataaagg ccggataaaa cttgtgctta
     6181 tttttcttta cggtctttaa aaaggccgta atatccagct gaacggtctg gttataggta
     6241 cattgagcaa ctgactgaaa tgcctcaaaa tgttctttac gatgccattg ggatatatca
     6301 acggtggtat atccagtgat ttttttctcc attttagctt ccttagctcc tgaaaatctc
     6361 gataactcaa aaaatacgcc cggtagtgat cttatttcat tatggtgaaa gttggaacct
     6421 cttacgtgcc gatcaacgtc tcattttcgc caaaagttgg cccagggctt cccggtatca
     6481 acagggacac caggatttat ttattctgcg aagtgatctt ccgtcacagg tatttattcg
     6541 gcgcaaagtg cgtcgggtga tgctgccaac ttactgattt agtgtatgat ggtgtttttg
     6601 aggtgctcca gtggcttctg tttctatcag ctgtccctcc tgttcagcta ctgacggggt
     6661 ggtgcgtaac ggcaaaagca ccgccggaca tcagcgctag cggagtgtat actggcttac
     6721 tatgttggca ctgatgaggg tgtcagtgaa gtgcttcatg tggcaggaga aaaaaggctg
     6781 caccggtgcg tcagcagaat atgtgataca ggatatattc cgcttcctcg ctcactgact
     6841 cgctacgctc ggtcgttcga ctgcggcgag cggaaatggc ttacgaacgg ggcggagatt
     6901 tcctggaaga tgccaggaag atacttaaca gggaagtgag agggccgcgg caaagccgtt
     6961 tttccatagg ctccgccccc ctgacaagca tcacgaaatc tgacgctcaa atcagtggtg
     7021 gcgaaacccg acaggactat aaagatacca ggcgtttccc ctggcggctc cctcgtgcgc
     7081 tctcctgttc ctgcctttcg gtttaccggt gtcattccgc tgttatggcc gcgtttgtct
     7141 cattccacgc ctgacactca gttccgggta ggcagttcgc tccaagctgg actgtatgca
     7201 cgaacccccc gttcagtccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     7261 cccggaaaga catgcaaaag caccactggc agcagccact ggtaattgat ttagaggagt
     7321 tagtcttgaa gtcatgcgcc ggttaaggct aaactgaaag gacaagtttt ggtgactgcg
     7381 ctcctccaag ccagttacct cggttcaaag agttggtagc tcagagaacc ttcgaaaaac
     7441 cgccctgcaa ggcggttttt tcgttttcag agcaagagat tacgcgcaga ccaaaacgat
     7501 ctcaagaaga tcatcttatt aatcagataa aatatttcta gatttcagtg caatttatct
     7561 cttcaaatgt agcacctgaa gtcagcccca tacgatataa gttgtaattc tcatgttagt
     7621 catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg gcatcggtcg
     7681 agatcccggt gcctaatgag tgagctaact tacattaatt gcgttgcgct cactgcccgc
     7741 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     7801 aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag acgggcaaca
     7861 gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc acgctggttt
     7921 gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa catgagctgt
     7981 cttcggtatc gtcgtatccc actaccgaga tgtccgcacc aacgcgcagc ccggactcgg
     8041 taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc gcagtgggaa
     8101 cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca ctccagtcgc
     8161 cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc cagccagcca
     8221 gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac
     8281 ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag aaaataatac
     8341 tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta gtgcaggcag
     8401 cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc ccactgacgc
     8461 gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt cgttctacca
     8521 tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc gcgacaattt
     8581 gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac gactgtttgc
     8641 ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc gccgcttcca
     8701 ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg gaaacggtct
     8761 gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc acattcacca
     8821 ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt
     8881 cgatggtgtc cgggatctcg acgctctccc ttatgcgact cctgcattag gaaattaata
     8941 cgactcacta ta
//
