LOCUS       sequence-494362-       10098 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses Myc-tagged ColH and His-tagged HylB as two separate 
            proteins in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10098)
  AUTHORS   Avsharian LC, Loganathan S, Ebelt ND, Shalamzari AF, Rodarte Munoz 
            I, Manuel ER
  TITLE     Tumor-Colonizing E. coli Expressing Both Collagenase and 
            Hyaluronidase Enhances Therapeutic Efficacy of Gemcitabine in 
            Pancreatic Cancer Models.
  JOURNAL   Biomolecules. 2024 Nov 17;14(11):1458. doi: 10.3390/biom14111458.
  PUBMED    39595636
REFERENCE   2  (bases 1 to 10098)
  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: 
            "Biomolecules."; date: "2024-11-17"; volume: "14(11):1458. doi"; 
            pages: " 10.3390/biom14111458"
COMMENT     Sequence Label: pBAD-ColH-HylB
FEATURES             Location/Qualifiers
     source          1..10098
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..285
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     208..227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             3388..3417
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             6399..6416
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6499..6516)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      6669..6755
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      6847..6874
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        6893..6984
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6985..7053,7054..7845)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(7203..7222)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8016..8604
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8505..8524
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8758..8775
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    8790..8930
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(8856..8875)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9016..9038
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(9194..10072)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
       61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
      121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
      181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg
      241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg ttttttgggc
      301 taacaggagg aattaaccat gggcatgaaa cgtaaatgcc tgtctaaacg tctgatgctg
      361 gcgatcacta tggcgaccat cttcaccgtt aactctaccc tgccgatcta cgcggcggtt
      421 gacaaaaaca acgcgaccgc ggcggttcag aacgaatcta aacgttacac cgtttcttac
      481 ctgaaaaccc tgaactacta cgacctggtt gacctgctgg ttaaaaccga aatcgaaaac
      541 ctgccggacc tgttccagta ctcttctgac gcgaaagaat tctacggcaa caaaacccgt
      601 atgtctttca tcatggacga aatcggccgt cgtgcgccgc agtacaccga aatcgaccac
      661 aaaggcatcc cgaccctggt tgaagttgtt cgtgcgggct tctacctggg cttccacaac
      721 aaagaactga acgaaatcaa caaacgttct ttcaaagaac gtgttatccc gtctatcctg
      781 gcgatccaga aaaacccgaa cttcaaactg ggcaccgaag ttcaggacaa aatcgtttct
      841 gcgaccggcc tgctggcggg caacgaaacc gcgccgccgg aagttgttaa caacttcacc
      901 ccgatcctgc aggactgcat caaaaacatc gaccgttacg cgctggacga cctgaaatct
      961 aaagcgctgt tcaacgttct ggcggcgccg acctacgaca tcaccgaata cctgcgtgcg
     1021 accaaagaaa aaccggaaaa caccccgtgg tacggcaaaa tcgacggctt catcaacgaa
     1081 ctgaaaaaac tggcgctgta cggcaaaatc aacgacaaca actcttggat catcgacaac
     1141 ggcatctacc acatcgcgcc gctgggcaaa ctgcactcta acaacaaaat cggcatcgaa
     1201 accctgaccg aagttatgaa agtttacccg tacctgtcta tgcagcacct gcagtctgcg
     1261 gaccagatca aacgtcacta cgactctaaa gacgcggaag gcaacaaaat cccgctggac
     1321 aaattcaaaa aagaaggcaa agaaaaatac tgcccgaaaa cctacacctt cgacgacggc
     1381 aaagttatca tcaaagcggg cgcgcgtgtt gaagaagaaa aagttaaacg tctgtactgg
     1441 gcgtctaaag aagttaactc tcagttcttc cgtgtttacg gcatcgacaa accgctggaa
     1501 gaaggcaacc cggacgacat cctgactatg gttatctaca actctccgga agaatacaaa
     1561 ctgaactctg ttctgtacgg ctacgacacc aacaacggcg gcatgtacat cgaaccggaa
     1621 ggcaccttct tcacctacga acgtgaagcg caggaatcta cctacaccct ggaagaactg
     1681 ttccgtcacg aatacaccca ctacctgcag ggccgttacg cggttccggg ccagtggggc
     1741 cgtaccaaac tgtacgacaa cgaccgtctg acctggtacg aagaaggcgg cgcggaactg
     1801 ttcgcgggct ctacccgtac ctctggcatc ctgccgcgta aatctatcgt ttctaacatc
     1861 cacaacacca cccgtaacaa ccgttacaaa ctgtctgaca ccgttcactc taaatacggc
     1921 gcgtctttcg aattctacaa ctacgcgtgc atgttcatgg actacatgta caacaaagac
     1981 atgggcatcc tgaacaaact gaacgacctg gcgaaaaaca acgacgttga cggctacgac
     2041 aactacatcc gtgacctgtc ttctaactac gcgctgaacg acaaatacca ggaccacatg
     2101 caggaacgta tcgacaacta cgaaaacctg accgttccgt tcgttgcgga cgactacctg
     2161 gttcgtcacg cgtacaaaaa cccgaacgaa atctactctg aaatctctga agttgcgaaa
     2221 ctgaaagacg cgaaatctga agttaaaaaa tctcagtact tctctacctt caccctgcgt
     2281 ggctcttaca ccggcggcgc gtctaaaggc aaactggaag accagaaagc gatgaacaaa
     2341 ttcatcgacg actctctgaa aaaactggac acctactctt ggtctggcta caaaaccctg
     2401 accgcgtact tcaccaacta caaagttgac tcttctaacc gtgttaccta cgacgttgtt
     2461 ttccacggct acctgccgaa cgaaggcgac tctaaaaact ctctgccgta cggcaaaatc
     2521 aacggcacct acaaaggcac cgaaaaagaa aaaatcaaat tctcttctga aggctctttc
     2581 gacccggacg gcaaaatcgt ttcttacgaa tgggacttcg gcgacggcaa caaatctaac
     2641 gaagaaaacc cggaacactc ttacgacaaa gttggcacct acaccgttaa actgaaagtt
     2701 accgacgaca aaggcgaatc ttctgtttct accaccaccg cggaaatcaa agacctgtct
     2761 gaaaacaaac tgccggttat ctacatgcac gttccgaaat ctggcgcgct gaaccagaaa
     2821 gttgttttct acggcaaagg cacctacgac ccggacggct ctatcgcggg ctaccagtgg
     2881 gacttcggcg acggctctga cttctcttct gaacagaacc cgtctcacgt ttacaccaaa
     2941 aaaggcgaat acaccgttac cctgcgtgtt atggactctt ctggccagat gtctgaaaaa
     3001 accatgaaaa tcaaaatcac cgacccggtt tacccgatcg gcaccgaaaa agaaccgaac
     3061 aactctaaag aaaccgcgtc tggcccgatc gttccgggca tcccggtttc tggcaccatc
     3121 gaaaacacct ctgaccagga ctacttctac ttcgacgtta tcaccccggg cgaagttaaa
     3181 atcgacatca acaaactggg ctacggcggc gcgacctggg ttgtttacga cgaaaacaac
     3241 aacgcggttt cttacgcgac cgacgacggc cagaacctgt ctggcaaatt caaagcggac
     3301 aaaccgggcc gttactacat ccacctgtac atgttcaacg gctcttacat gccgtaccgt
     3361 atcaacatcg aaggctctgt tggccgtgaa cagaaactga tctctgaaga agacctgtaa
     3421 tagtgattta agaaggagat atacatatga aacaggttgt tgacaaccag acccagaaca
     3481 aagaactggt taaaaacggc gacttcaacc agaccaaccc ggtttctggc tcttggtctc
     3541 acacctctgc gcgtgaatgg tctgcgtgga tcgacaaaga aaacaccgcg gacaaatctc
     3601 cgatcatcca gcgtaccgaa cagggccagg tttctctgtc ttctgacaaa ggcttccgtg
     3661 gcgcggttac ccagaaagtt aacatcgacc cgaccaaaaa atacgaagtt aaattcgaca
     3721 tcgaaacctc taacaaagcg ggccaggcgt tcctgcgtat catggaaaaa aaagacaaca
     3781 acacccgtct gtggctgtct gaaatgacct ctggcaccac caacaaacac accctgacca
     3841 aaatctacaa cccgaaactg aacgtttctg aagttaccct ggaactgtac tacgaaaaag
     3901 gcaccggctc tgcgaccttc gacaacatct ctatgaaagc gaaaggcccg aaagactctg
     3961 aacacccgca gccggttacc acccagatcg aagaatctgt taacaccgcg ctgaacaaaa
     4021 actacgtttt caacaaagcg gactaccagt acaccctgac caacccgtct ctgggcaaaa
     4081 tcgttggcgg catcctgtac ccgaacgcga ccggctctac caccgttaaa atctctgaca
     4141 aatctggcaa aatcatcaaa gaagttccgc tgtctgttac cgcgtctacc gaagacaaat
     4201 tcaccaaact gctggacaaa tggaacgacg ttaccatcgg caaccacgtt tacgacacca
     4261 acgactctaa catgcagaaa atcaaccaga aactggacga aaccaacgcg aaaaacatca
     4321 aaaccatcaa actggactct aaccacacct tcctgtggaa agacctggac aacctgaaca
     4381 actctgcgca gctgaccgcg acctaccgtc gtctggaaga cctggcgaaa cagatcacca
     4441 acccgcactc taccatctac aaaaacgaaa aagcgatccg taccgttaaa gaatctctgg
     4501 cgtggctgca ccagaacttc tacaacgtta acaaagacat cgaaggctct gcgaactggt
     4561 gggacttcga aatcggcgtt ccgcgttcta tcaccgcgac cctggcgctg atgaacaact
     4621 acttcaccga cgcggaaatc aaaacctaca ccgacccgat cgaacacttc gttccggacg
     4681 cgggctactt ccgtaaaacc ctggacaacc cgttcaaagc gctgggcggc aacctggttg
     4741 acatgggccg tgttaaaatc atcgaaggcc tgctgcgtaa agacaacacc atcatcgaaa
     4801 aaacctctca ctctctgaaa aacctgttca ccaccgcgac caaagcggaa ggcttctacg
     4861 cggacggctc ttacatcgac cacaccaacg ttgcgtacac cggcgcgtac ggcaacgttc
     4921 tgatcgacgg cctgacccag ctgctgccga tcatccagga aaccgactac aaaatctcta
     4981 accaggaact ggacatggtt tacaaatgga tcaaccagtc tttcctgccg ctgatcgtta
     5041 aaggcgaact gatggacatg tctcgtggcc gttctatctc tcgtgaagcg gcgtcttctc
     5101 acgcggcggc ggttgaagtt ctgcgtggct tcctgcgtct ggcgaacatg tctaacgaag
     5161 aacgtaacct ggacctgaaa tctaccatca aaaccatcat cacctctaac aaattctaca
     5221 acgttttcaa caacctgaaa tcttactctg acatcgcgaa catgaacaaa atgctgaacg
     5281 actctaccgt tgcgaccaaa ccgctgaaat ctaacctgtc taccttcaac tctatggacc
     5341 gtctggcgta ctacaacgcg gaaaaagact tcggcttcgc gctgtctctg cactctaaac
     5401 gtaccctgaa ctacgaaggc atgaacgacg aaaacacccg tgactggtac accggcgacg
     5461 gcatgttcta cctgtacaac tctgaccagt ctcactactc taaccacttc tggccgaccg
     5521 ttaacccgta caaaatggcg ggcaccaccg aaaaagacgc gaaacgtgaa gacaccacca
     5581 aagaatttat gtctaaacac tctaaagacg cgaaagaaaa aaccggccag gttaccggca
     5641 cctctgactt cgttggctct gttaaactga acgaccactt cgcgctggcg gcgatggact
     5701 tcaccaactg ggaccgtacc ctgaccgcgc agaaaggctg ggttatcctg aacgacaaaa
     5761 tcgttttcct gggctctaac atcaaaaaca ccaacggcat cggcaacgtt tctaccacca
     5821 tcgaccagcg taaagacgac tctaaaaccc cgtacaccac ctacgttaac ggcaaaacca
     5881 tcgacctgaa acaggcgtct tctcagcagt tcaccgacac caaatctgtt ttcctggaat
     5941 ctaaagaacc gggccgtaac atcggctaca tcttcttcaa aaactctacc atcgacatcg
     6001 aacgtaaaga acagaccggc acctggaact ctatcaaccg tacctctaaa aacacctcta
     6061 tcgtttctaa cccgttcatc accatctctc agaaacacga caacaaaggc gactcttacg
     6121 gctacatgat ggttccgaac atcgaccgta cctctttcga caaactggcg aactctaaag
     6181 aagttgaact gctggaaaac tcttctaaac agcaggttat ctacgacaaa aactctcaga
     6241 cctgggcggt tatcaaacac gacaaccagg aatctctgat caacaaccag ttcaaaatga
     6301 acaaagcggg cctgtacctg gttcagaaag ttggcaacga ctaccagaac gtttactacc
     6361 agccgcagac catgaccaaa accgaccagc tggcgatcca tcaccaccac catcactgat
     6421 agtaactcga gatctgcagc tggtaccata tgggaattcg aagcttggct gttttggcgg
     6481 atgagagaag attttcagcc tgatacagat taaatcagaa cgcagaagcg gtctgataaa
     6541 acagaatttg cctggcggca gtagcgcggt ggtcccacct gaccccatgc cgaactcaga
     6601 agtgaaacgc cgtagcgccg atggtagtgt ggggtctccc catgcgagag tagggaactg
     6661 ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt
     6721 gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gggagcggat ttgaacgttg
     6781 cgaagcaacg gcccggaggg tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa
     6841 ttaagcagaa ggccatcctg acggatggcc tttttgcgtt tctacaaact cttttgttta
     6901 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     6961 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     7021 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     7081 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     7141 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     7201 ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc
     7261 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     7321 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     7381 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     7441 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     7501 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     7561 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     7621 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     7681 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     7741 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     7801 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     7861 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     7921 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     7981 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     8041 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     8101 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     8161 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     8221 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     8281 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     8341 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     8401 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     8461 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     8521 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     8581 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     8641 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     8701 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     8761 gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg
     8821 tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga tgccgcatag
     8881 ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg ccccgacacc
     8941 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     9001 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     9061 gcgcgaggca gcagatcaat tcgcgcgcga aggcgaagcg gcatgcataa tgtgcctgtc
     9121 aaatggacga agcagggatt ctgcaaaccc tatgctactc cgtcaagccg tcaattgtct
     9181 gattcgttac caattatgac aacttgacgg ctacatcatt cactttttct tcacaaccgg
     9241 cacggaactc gctcgggctg gccccggtgc attttttaaa tacccgcgag aaatagagtt
     9301 gatcgtcaaa accaacattg cgaccgacgg tggcgatagg catccgggtg gtgctcaaaa
     9361 gcagcttcgc ctggctgata cgttggtcct cgcgccagct taagacgcta atccctaact
     9421 gctggcggaa aagatgtgac agacgcgacg gcgacaagca aacatgctgt gcgacgctgg
     9481 cgatatcaaa attgctgtct gccaggtgat cgctgatgta ctgacaagcc tcgcgtaccc
     9541 gattatccat cggtggatgg agcgactcgt taatcgcttc catgcgccgc agtaacaatt
     9601 gctcaagcag atttatcgcc agcagctccg aatagcgccc ttccccttgc ccggcgttaa
     9661 tgatttgccc aaacaggtcg ctgaaatgcg gctggtgcgc ttcatccggg cgaaagaacc
     9721 ccgtattggc aaatattgac ggccagttaa gccattcatg ccagtaggcg cgcggacgaa
     9781 agtaaaccca ctggtgatac cattcgcgag cctccggatg acgaccgtag tgatgaatct
     9841 ctcctggcgg gaacagcaaa atatcacccg gtcggcaaac aaattctcgt ccctgatttt
     9901 tcaccacccc ctgaccgcga atggtgagat tgagaatata acctttcatt cccagcggtc
     9961 ggtcgataaa aaaatcgaga taaccgttgg cctcaatcgg cgttaaaccc gccaccagat
    10021 gggcattaaa cgagtatccc ggcagcaggg gatcattttg cgcttcagcc atacttttca
    10081 tactcccgcc attcagag
//
