LOCUS       sequence-233743-        8954 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 8954)
  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 8954)
  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..8954
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..17
                     /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(2489..3022)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3211..3216,3217..3233,3234..3239))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     oriT            3253..3361
                     /note="incP origin of transfer"
     protein_bind    3501..3548
                     /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             4289..4333
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(4371..4389)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4385..4432
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(4653..5312)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     5227..5246
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(5313..5415)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(5807..5824)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5941..6486)
                     /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(6021..6040)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6711..7793)
                     /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     7755..7774
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(7794..7871)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     7917..7936
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7917..7935
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    7936..7960
                     /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             8016..8033
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 ttgtgagcgg ataacaattt ccacccacag ccagcctttt ttaggcgcca atctaggcac
       61 aggaatacgg gtgaggcggc ttccggatag ttgctggggc tgagcgggac atatgacgag
      121 catagggcct aagtcattac ttaggagcct cgaaatggca ctctctgatc tgatcgtccg
      181 gcaggccaag accaccggca aacgctacac cctctacgac aacgactgcc tgagcctgat
      241 ggtctctgcc gcaggcggca agtcttggat gttccgctac tcctggctgg gcaagcaaaa
      301 gcgcatggcc ctgggcggct atcctgccct cagcctgcgc gaggcccgcg ccgagcggga
      361 caaggcccag gccctgatcg ccagggggat cgatccccaa atcgaacgcg atcagcgtcg
      421 gcacgcggcc aagctggccg gcgaatacac cttcaagaac gtcttcgatg cctgggtcga
      481 gcatcgccgc aaggaactca aggaaggccg ccagagtacg ctttcgcaga tcctgcgcat
      541 cttcaacaag gacgtgctgc ccaccctagg gaagatgtcc atctatgaca ttcgccggcc
      601 ccagctcctg ggcgtcctgg cggcgatcga gaagcgcaag gcgttcacca cggccgagaa
      661 ggtccgcacc tggttcaacc agatgttccg ctatgccttg gtcatcgccg aggggctgga
      721 agtcaacccg gctgcggacc tggacgtggt ggccgagccc aagcccccgg tggcccacaa
      781 cccctatctg cacctgcccg aactgcccga gttccttcag aagctgcgac gctacaaccc
      841 ccggggctgg cagacccagc ttggcgtgcg gctactgttc ctgacggggg tgcgcacggg
      901 cgagctgcgg ttggccgaac ccgagcagtt cgacctcgac cggggctttt ggatcatccc
      961 gccggaggtc gtcaagcaac tccaggacga aatgcgcaag gccggcaagc ggccgcagga
     1021 cgtgccgccc tacatcgtgc ccctgtccct gcaggccatc gagatcgtgc gctatctcct
     1081 gggcgtgatg cggccggcgc agaagtacct gctgtcgcac cgcagcgaac tcaagaaacg
     1141 catcagcgag aacaccctca acaaggccgt gcagctcatg ggctacgagg ggcgcctgac
     1201 cgggcacggc atccgcggca ccatctcgac ggcactcaac gagatcggct acccgaagat
     1261 ttgggtggac gcgcagcttt cgcactccga ccccaacaag gtgagttcgg cctacaacca
     1321 cgccaagtac gtggagccgc gtcgccgcat gatgcaggac tgggccgatc gcctggacct
     1381 gctcgaacag ggcgaagtgc aagctgcgag cgcgcacctg accatccgta tcgacggggt
     1441 gccagcgatg gcggaagtgg aggaagcggt ggacgtggcc ccgacggtcg ccgagcccgg
     1501 tgtctccggc tcaccgcccg tggcggccac gcccatcgtc gtgaccccca acagcggcgg
     1561 aatcacgttc cagcgcctgt cgcaggtgcc gccgcctccg gcgcatgctc ctgagtcgga
     1621 ggtctctgcc atccagcgcg agcgtgagga aatgctggcc atgtacgagt cgccgaacaa
     1681 tctgccggtc gcgctgttcg ggaagctggc tgggaagtcc aaagaccaga tcaaccgcga
     1741 gctgaaggca ggcaagctac tgtccatcag cttgggcaac cgggggcagc gcgttcccga
     1801 ctggcaactg gtgccgctca agcgccggct ggcccaagcg ctcatgaacc aatgcccgca
     1861 tgtagattct tgggcgctgt atcgtctgtt gaccaagccc cattccaacc tcggaaaccg
     1921 tgcagccatc gatgtcgtca caccgaccaa cgtgggcaaa gtattgcaag ccgtcacgcc
     1981 ctataaggag ttcgagcgct ccagtacaga cgagacaccc cagttctccg agttcgttcg
     2041 ccagttgcaa cgccacgtga atgcgctcga agaagcgccg tgctgacgag ccaggacttc
     2101 gacgccaacg gcgtgccgca acgcgtgttt acctagacgc taggagtgcg ttaaatcccg
     2161 tcgtgggcgg ctattgcaac gaactgatca accagacggc caagaccgac tgcaattcct
     2221 cccggtaagg tttttcgcct ccgcgtccgg atgtccgttc cttgagcggc gggtaggagg
     2281 cctgtaccca ggatttatga ggttccccgc cccaaggtcc attcccatgt gcgaaccgac
     2341 agctcggcaa acagacctgc ctgtacgtag gggtcgtggg ccacgaaggc gtgcacgtcg
     2401 tcgatatgcg ccgcctgcac aagcggcgtt gtgacaattt accgaacaac tccgcggccg
     2461 ggaagccgat ctcggcttga acgaattgtt aggtggcggt acttgggtcg atatcaaagt
     2521 gcatcacttc ttcccgtatg cccaactttg tatagagagc cactgcggga tcgtcaccgt
     2581 aatctgcttg cacgtagatc acataagcac caagcgcgtt ggcctcatgc ttgaggagat
     2641 tgatgagcgc ggtggcaatg ccctgcctcc ggtgctcgcc ggagactgcg agatcataga
     2701 tatagatctc actacgcggc tgctcaaact tgggcagaac gtaagccgcg agagcgccaa
     2761 caaccgcttc ttggtcgaag gcagcaagcg cgatgaatgt cttactacgg agcaagttcc
     2821 cgaggtaatc ggagtccggc tgatgttggg agtaggtggc tacgtctccg aactcacgac
     2881 cgaaaagatc aagagcagcc cgcatggatt tgacttggtc agggccgagc ctacatgtgc
     2941 gaatgatgcc catacttgag ccacctaact ttgttttagg gcgactgccc tgctgcgtaa
     3001 catcgttgct gctgcgtaac atcgttgctg ctccataaca tcaaacatcg acccacggcg
     3061 taacgcgctt gctgcttgga tgcccgaggc atagactgta caaaaaaaca gtcataacaa
     3121 gccatgaaaa ccgccactgc gccgttacca ccgctgcgtt cggtcaaggt tctggaccag
     3181 ttgcgtgagc gcatacgcta cttgcattac agtttacgaa ccgaacaggc ttatgtcaat
     3241 tcggccggcc tacggccagc ctcgcagagc aggattcccg ttgagcaccg ccaggtgcga
     3301 ataagggaca gtgaagaagg aacacccgct cgcgggtggg cctacttcac ctatcctgcc
     3361 cggctgacgc cgttggatac accaaggaaa gtctacacga accctttggc aaaatcctgt
     3421 atatcgtgcg aaaaaggatg gatataccga aaaaatcgct ataatgaccc cgaagcaggg
     3481 ttatgcagcg gaaaagaaaa gaagttccta ttccgaagtt cctattctct agaaagtata
     3541 ggaacttcat gcatgatctg aagatcagca gttcaacctg ttgatagtac gtactaagct
     3601 ctcatgtttc acgtactaag ctctcatgtt taacgtacta agctctcatg tttaacgaac
     3661 taaaccctca tggctaacgc tccatcgtat ccgtattcct cttatttttt ctctttctct
     3721 atatccattt ctttctctct aggtgtgtcc tctctctctc ttcaatttct ctactccgca
     3781 ttccaacgca tccttccccc aacctcccat ttcctcctta cggcccgata gcgatcgtct
     3841 ttccctcgct atcactcgct accggcccct cctctgcacc gtaacctcct acgtatttac
     3901 catatcataa agttttttcc gacgcttatc gctgaccccc tgtcgccctc ctattggctt
     3961 ccggattatc ttcttgtcca taaggtgatc catgcttcct gaagattccc gaaatgtgtc
     4021 cactttggcg gggaatcatt ccatccactt ctttctctct cgctttcctc attcggcgct
     4081 ccccttccgc gtctcattgg tcttccgctc cgtttttgct ttgccgatgt tacttgggga
     4141 gaggtgcgat aatcctttcg caaaaactcg gtttgacgcc tcccatggta taaatagtgg
     4201 gcgtttggta cggcggctcc gccacggcat caatcggccc aatcggcaaa tttgcggtca
     4261 atcgtcttgc gcgaaactcg agtctggtaa agaaaccgct gctgcgaaat ttgaacgcca
     4321 gcacatggac tcgtctacta gcgcagctta attaacctag gctgctgcca ccgctgagca
     4381 ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaacc
     4441 tcaggcattt gagaagcaca cggtcacact gcttccggta gtcaataaac cggtaaacca
     4501 gcaatagaca taagcggcta tttaacgacc ctgccctgaa ccgacgaccg ggtcgaattt
     4561 gctttcgaat ttctgccatt catccgctta ttatcactta ttcaggcgta gcaaccaggc
     4621 gtttaagggc accaataact gccttaaaaa aattacgccc cgccctgcca ctcatcgcag
     4681 tactgttgta attcattaag cattctgccg acatggaagc catcacaaac ggcatgatga
     4741 acctgaatcg ccagcggcat cagcaccttg tcgccttgcg tataatattt gcccatagtg
     4801 aaaacggggg cgaagaagtt gtccatattg gccacgttta aatcaaaact ggtgaaactc
     4861 acccagggat tggctgagac gaaaaacata ttctcaataa accctttagg gaaataggcc
     4921 aggttttcac cgtaacacgc cacatcttgc gaatatatgt gtagaaactg ccggaaatcg
     4981 tcgtggtatt cactccagag cgatgaaaac gtttcagttt gctcatggaa aacggtgtaa
     5041 caagggtgaa cactatccca tatcaccagc tcaccgtctt tcattgccat acggaactcc
     5101 ggatgagcat tcatcaggcg ggcaagaatg tgaataaagg ccggataaaa cttgtgctta
     5161 tttttcttta cggtctttaa aaaggccgta atatccagct gaacggtctg gttataggta
     5221 cattgagcaa ctgactgaaa tgcctcaaaa tgttctttac gatgccattg ggatatatca
     5281 acggtggtat atccagtgat ttttttctcc attttagctt ccttagctcc tgaaaatctc
     5341 gataactcaa aaaatacgcc cggtagtgat cttatttcat tatggtgaaa gttggaacct
     5401 cttacgtgcc gatcaacgtc tcattttcgc caaaagttgg cccagggctt cccggtatca
     5461 acagggacac caggatttat ttattctgcg aagtgatctt ccgtcacagg tatttattcg
     5521 gcgcaaagtg cgtcgggtga tgctgccaac ttactgattt agtgtatgat ggtgtttttg
     5581 aggtgctcca gtggcttctg tttctatcag ctgtccctcc tgttcagcta ctgacggggt
     5641 ggtgcgtaac ggcaaaagca ccgccggaca tcagcgctag cggagtgtat actggcttac
     5701 tatgttggca ctgatgaggg tgtcagtgaa gtgcttcatg tggcaggaga aaaaaggctg
     5761 caccggtgcg tcagcagaat atgtgataca ggatatattc cgcttcctcg ctcactgact
     5821 cgctacgctc ggtcgttcga ctgcggcgag cggaaatggc ttacgaacgg ggcggagatt
     5881 tcctggaaga tgccaggaag atacttaaca gggaagtgag agggccgcgg caaagccgtt
     5941 tttccatagg ctccgccccc ctgacaagca tcacgaaatc tgacgctcaa atcagtggtg
     6001 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggcggct ccctcgtgcg
     6061 ctctcctgtt cctgcctttc ggtttaccgg tgtcattccg ctgttatggc cgcgtttgtc
     6121 tcattccacg cctgacactc agttccgggt aggcagttcg ctccaagctg gactgtatgc
     6181 acgaaccccc cgttcagtcc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     6241 acccggaaag acatgcaaaa gcaccactgg cagcagccac tggtaattga tttagaggag
     6301 ttagtcttga agtcatgcgc cggttaaggc taaactgaaa ggacaagttt tggtgactgc
     6361 gctcctccaa gccagttacc tcggttcaaa gagttggtag ctcagagaac cttcgaaaaa
     6421 ccgccctgca aggcggtttt ttcgttttca gagcaagaga ttacgcgcag accaaaacga
     6481 tctcaagaag atcatcttat taatcagata aaatatttct agatttcagt gcaatttatc
     6541 tcttcaaatg tagcacctga agtcagcccc atacgatata agttgtaatt ctcatgttag
     6601 tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag ggcatcggtc
     6661 gagatcccgg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc tcactgcccg
     6721 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     6781 gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga gacgggcaac
     6841 agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc cacgctggtt
     6901 tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata acatgagctg
     6961 tcttcggtat cgtcgtatcc cactaccgag atgtccgcac caacgcgcag cccggactcg
     7021 gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga
     7081 acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg
     7141 ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc
     7201 agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga
     7261 cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga gaaaataata
     7321 ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt agtgcaggca
     7381 gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag cccactgacg
     7441 cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct tcgttctacc
     7501 atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc cgcgacaatt
     7561 tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa cgactgtttg
     7621 cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat cgccgcttcc
     7681 actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc
     7741 tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt cacattcacc
     7801 accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt tttgcgccat
     7861 tcgatggtgt ccgggatctc gacgctctcc cttatgcgac tcctgcatta ggaaattaat
     7921 acgactcact ataggggaat tgtgagcgga taacaattcc cctgtagaaa taattttgtt
     7981 taactttaat aaggagatat accatgggca gcagccatca ccatcatcac cacagccagg
     8041 atccgaattc gagctcggcg cgcctgcagg tcgacaagct ttctatcgcc ttcttgacga
     8101 gttcttctga ggttaactgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     8161 cccgaaaagt gccacctggg tccttttcat cacgtgctat aaaaataatt ataatttaaa
     8221 ttttttaata taaatatata aattaaaaat agaaagtaaa aaaagaaatt aaagaaaaaa
     8281 tagtttttgt tttccgaaga tgtaaaagac tctaggggga tcgccaacaa atactacctt
     8341 ttatcttgct cttcctgctc tcaggtatta atgccgaatt gtttcatctt gtctgtgtag
     8401 aagaccacac acgaaaatcc tgtgatttta cattttactt atcgttaatc gaatgtatat
     8461 ctatttaatc tgcttttctt gtctaataaa tatatatgta aagtacgctt tttgttgaaa
     8521 ttttttaaac ctttgtttat ttttttttct tcattccgta actcttctac cttctttatt
     8581 tactttctaa aatccaaata caaaacataa aaataaataa acacagagta aattcccaaa
     8641 ttattccatc attaaaagat acgaggcgcg tgtaagttac aggcaagcgg ccgaattctg
     8701 ctgggtgtgg gtctatggat gccagtggaa ctctttggac gatctttgtt ggcggggacg
     8761 aaaaaatagg ctacaatcta ggtcttcgct ggccacctcg gcgaagaaag agcttgggaa
     8821 tcaacgactt actgccctag taaacgtctc gtttcccgct ccaagttatg atctacagac
     8881 ttctactgac gtctgtaagt cgttgaaaag aggggcttcg gccccttttt tcattccagc
     8941 gagcgccacg gaaa
//
