LOCUS       sequence-264461-        5456 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For L-arabinose inducible bacterial expression of S. typhimurium 
            codon-optimized hyaluronidase (Streptomyces koganeinsis UniProt 
            A0A0U2E2) with N-terminal 6x His tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5456)
  AUTHORS   Ebelt ND, Zuniga E, Passi KB, Sobocinski LJ, Manuel ER
  TITLE     Hyaluronidase-Expressing Salmonella Effectively Targets 
            Tumor-Associated Hyaluronic Acid in Pancreatic Ductal 
            Adenocarcinoma.
  JOURNAL   Mol Cancer Ther. 2019 Nov 6. pii: 1535-7163.MCT-19-0556. doi: 
            10.1158/1535-7163.MCT-19-0556.
  PUBMED    31694889
REFERENCE   2  (bases 1 to 5456)
  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: "Mol Cancer 
            Ther. 2019 Nov 6. pii: 1535-7163.MCT-19-0556. doi: 
            10.1158/1535-7163.MCT-19-0556."
COMMENT     Sequence Label: pBAD-bHs
FEATURES             Location/Qualifiers
     source          1..5456
                     /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"
     RBS             312..334
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             354..371
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             375..407
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             420..440
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(1239..1256)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1409..1495
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1587..1614
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1633..1724
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1725..1793,1794..2585)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1943..1962)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2627..3082
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2714..2733)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2924..2945
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      3193..3781
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3682..3701
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3935..3952
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3967..4107
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4033..4052)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4193..4215
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4353..4372
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(4552..5430)
                     /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 atactacctg acgcttttta tcgcaactct ctactgtttc tccatacccg tttttttggg
      301 ctagaaataa ttttgtttaa ctttaagaag gagatataca tatgcggggt tctcatcatc
      361 atcatcatca tggtatggct agcatgactg gtggacagca aatgggtcgg gatctgtacg
      421 agaacctgta cttccagggc tcgagcatgc cggttgcgcg tcgtctgttc ctgggctctt
      481 tcaccgcggg cgcggttacc gttgcgaccg cggcggcgac cggcaccgcg tctgcggcgg
      541 gcgaaaacgg cgcgaccacc accttcgacg gcccggttgc ggcggaacgt ttctctgcgg
      601 acaccaccct ggaagcggcg ttcctgaaaa ccacctctga aaccaaccac gcggcgacca
      661 tctaccaggc gggcacctct ggcgacggcg cggcgctgaa cgttatctct gacaacccgg
      721 gcacctctgc gatgtacctg tctggcaccg aaaccgcgcg tggcaccctg aaaatcaccc
      781 accgtggcta cgcggacggc tctgacaaag acgcggcggc gctgtctctg gacctgcgtg
      841 ttgcgggcac cgcggcgcag ggcatctacg ttaccgcgac caacggcccg accaaaggca
      901 acctgatcgc gctgcgtaac aacaccggcc tggacgactt cgttgttaaa ggcaccggcc
      961 gtatcggcgt tggcatcgac cgtgcggcga ccccgcgtgc gcaggttcac atcgttcagc
     1021 gtggcgacgc gctggcggcg ctgctggttg aaggctctgt tcgtatcggc aacgcggcga
     1081 ccgttccgac ctctgttgac tcttctggcg gcggcgcgct gtacgcgtct ggcggcgcgc
     1141 tgctgtggcg tggctctaac ggcaccgtta ccaccatcgc gccggcgtaa tgagaattcg
     1201 aagcttggct gttttggcgg atgagagaag attttcagcc tgatacagat taaatcagaa
     1261 cgcagaagcg gtctgataaa acagaatttg cctggcggca gtagcgcggt ggtcccacct
     1321 gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt ggggtctccc
     1381 catgcgagag tagggaactg ccaggcatca aataaaacga aaggctcagt cgaaagactg
     1441 ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc
     1501 gggagcggat ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag gacgcccgcc
     1561 ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg acggatggcc tttttgcgtt
     1621 tctacaaact cttttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     1681 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     1741 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     1801 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     1861 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     1921 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc
     1981 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     2041 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     2101 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     2161 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     2221 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgca gcaatggcaa
     2281 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     2341 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     2401 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     2461 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     2521 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     2581 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttacgc gccctgtagc
     2641 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     2701 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     2761 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     2821 ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag
     2881 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     2941 actggaacaa cactcaaccc tatctcgggc tattcttttg atttataagg gattttgccg
     3001 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     3061 aaaatattaa cgtttacaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     3121 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     3181 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     3241 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     3301 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     3361 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     3421 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     3481 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     3541 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     3601 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     3661 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     3721 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     3781 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     3841 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     3901 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     3961 agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatctg
     4021 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta
     4081 tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc
     4141 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     4201 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagaa ggagatggcg
     4261 cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg
     4321 agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca
     4381 accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatctgctca
     4441 tgtttgacag cttatcatcg atgcataatg tgcctgtcaa atggacgaag cagggattct
     4501 gcaaacccta tgctactccg tcaagccgtc aattgtctga ttcgttacca attatgacaa
     4561 cttgacggct acatcattca ctttttcttc acaaccggca cggaactcgc tcgggctggc
     4621 cccggtgcat tttttaaata cccgcgagaa atagagttga tcgtcaaaac caacattgcg
     4681 accgacggtg gcgataggca tccgggtggt gctcaaaagc agcttcgcct ggctgatacg
     4741 ttggtcctcg cgccagctta agacgctaat ccctaactgc tggcggaaaa gatgtgacag
     4801 acgcgacggc gacaagcaaa catgctgtgc gacgctggcg atatcaaaat tgctgtctgc
     4861 caggtgatcg ctgatgtact gacaagcctc gcgtacccga ttatccatcg gtggatggag
     4921 cgactcgtta atcgcttcca tgcgccgcag taacaattgc tcaagcagat ttatcgccag
     4981 cagctccgaa tagcgccctt ccccttgccc ggcgttaatg atttgcccaa acaggtcgct
     5041 gaaatgcggc tggtgcgctt catccgggcg aaagaacccc gtattggcaa atattgacgg
     5101 ccagttaagc cattcatgcc agtaggcgcg cggacgaaag taaacccact ggtgatacca
     5161 ttcgcgagcc tccggatgac gaccgtagtg atgaatctct cctggcggga acagcaaaat
     5221 atcacccggt cggcaaacaa attctcgtcc ctgatttttc accaccccct gaccgcgaat
     5281 ggtgagattg agaatataac ctttcattcc cagcggtcgg tcgataaaaa aatcgagata
     5341 accgttggcc tcaatcggcg ttaaacccgc caccagatgg gcattaaacg agtatcccgg
     5401 cagcagggga tcattttgcg cttcagccat acttttcata ctcccgccat tcagag
//
