LOCUS       sequence-354970-        7266 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of bacterial PNGase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7266)
  TITLE     Bacterial expression for GST-His8-PNGase
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7266)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pHN767
FEATURES             Location/Qualifiers
     source          1..7266
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..47)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        115..219
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(220..288,289..1080)
                     /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(438..457)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1251..1839
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1740..1759
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1993..2010
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2025..2164
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2090..2109)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2250..2272
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2266..2457)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3769..4851)
                     /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     4813..4832
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4852..4929)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5135..5154
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5238..5257
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5238..5256
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5257..5281
                     /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)."
     RBS             5296..5318
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5325..5978
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     CDS             5991..6014
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     primer_bind     complement(7004..7022)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      7018..7065
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(7240..7245,7246..7262,7263..7266,1..2))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
ORIGIN
        1 aaacatgaga attcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat
       61 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
      121 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
      181 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
      241 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
      301 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
      361 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
      421 agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag
      481 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
      541 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
      601 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
      661 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
      721 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg
      781 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
      841 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
      901 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
      961 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     1021 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     1081 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     1141 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     1201 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     1261 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     1321 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     1381 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     1441 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     1501 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     1561 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     1621 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     1681 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     1741 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     1801 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     1861 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     1921 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     1981 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt
     2041 ctccttacgc atctgtgcgg tatttcacac cgcaatggtg cactctcagt acaatctgct
     2101 ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc
     2161 tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     2221 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     2281 gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga
     2341 ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa
     2401 tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga
     2461 tgcctccgtg taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag
     2521 gatgctcacg atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg
     2581 taaacaactg gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca
     2641 gcgcttcgtt aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca
     2701 gatccggaac ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg
     2761 gaaaccgaag accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct
     2821 tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc
     2881 tagccgggtc ctcaacgaca ggagcacgat catgcgcacc cgtggccagg acccaacgct
     2941 gcccgagatg cgccgcgtgc ggctgctgga gatggcggac gcgatggata tgttctgcca
     3001 agggttggtt tgcgcattca cagttctccg caagaattga ttggctccaa ttcttggagt
     3061 ggtgaatccg ttagcgaggt gccgccggct tccattcagg tcgaggtggc ccggctccat
     3121 gcaccgcgac gcaacgcggg gaggcagaca aggtataggg cggcgcctac aatccatgcc
     3181 aacccgttcc atgtgctcgc cgaggcggca taaatcgccg tgacgatcag cggtccaatg
     3241 atcgaagtta ggctggtaag agccgcgagc gatccttgaa gctgtccctg atggtcgtca
     3301 tctacctgcc tggacagcat ggcctgcaac gcgggcatcc cgatgccgcc ggaagcgaga
     3361 agaatcataa tggggaaggc catccagcct cgcgtcgcga acgccagcaa gacgtagccc
     3421 agcgcgtcgg ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg tttggtggcg
     3481 ggaccagtga cgaaggcttg agcgagggcg tgcaagattc cgaataccgc aagcgacagg
     3541 ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc
     3601 ggcacctgtc ctacgagttg catgataaag aagacagtca taagtgcggc gacgatagtc
     3661 atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga
     3721 gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct
     3781 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     3841 ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag
     3901 ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg
     3961 ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc
     4021 ttcggtatcg tcgtatccca ctaccgagat atccgcacca acgcgcagcc cggactcggt
     4081 aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac
     4141 gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc
     4201 ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag
     4261 acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc
     4321 caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact
     4381 gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc
     4441 ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg
     4501 ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat
     4561 cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg
     4621 cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc
     4681 cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac
     4741 tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg
     4801 ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac
     4861 cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc
     4921 gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg aagcagccca
     4981 gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg
     5041 cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca
     5101 tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag
     5161 caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga
     5221 tctcgatccc gcgaaattaa tacgactcac tataggggaa ttgtgagcgg ataacaattc
     5281 ccctctagaa ataattttgt ttaactttaa gaaggagata taccatgtcc cctatactag
     5341 gttattggaa aattaagggc cttgtgcaac ccactcgact tcttttggaa tatcttgaag
     5401 aaaaatatga agagcatttg tatgagcgcg atgaaggtga taaatggcga aacaaaaagt
     5461 ttgaattggg tttggagttt cccaatcttc cttattatat tgatggtgat gttaaattaa
     5521 cacagtctat ggccatcata cgttatatag ctgacaagca caacatgttg ggtggttgtc
     5581 caaaagagcg tgcagagatt tcaatgcttg aaggagcggt tttggatatt agatacggtg
     5641 tttcgagaat tgcatatagt aaagactttg aaactctcaa agttgatttt cttagcaagc
     5701 tacctgaaat gctgaaaatg ttcgaagatc gtttatgtca taaaacatat ttaaatggtg
     5761 atcatgtaac ccatcctgac ttcatgttgt atgacgctct tgatgttgtt ttatacatgg
     5821 acccaatgtg cctggatgcg ttcccaaaat tagtttgttt taaaaaacgt attgaagcta
     5881 tcccacaaat tgataagtac ttgaaatcca gcaagtatat agcatggcct ttgcagggct
     5941 ggcaagccac gtttggtggt ggcgaccatc ctccaaaatc ggatctgatg caccaccacc
     6001 accatcatca tcatgctccg gcagataata ccgtaaatat taaaacattc gacaaagtaa
     6061 aaaatgcctt tggtgacgga ttgtcccaaa gtgcggaagg aacctttaca tttccggccg
     6121 atgtaacagc cgtaaaaacg attaagatgt tcattaaaaa tgaatgtcct aataaaactt
     6181 gtgatgaatg ggatcgttat gccaatgttt atgtaaaaaa taaaacaaca ggtgagtggt
     6241 acgaaatagg acgctttatt actccatatt gggtgggaac ggaaaaatta cctcgtggac
     6301 tggaaattga tgttacagat ttcaaatctt tactatccgg aaatacagaa cttaaaattt
     6361 atacggagac atggctggcc aaaggaagag aatacagtgt agatttcgat attgtatacg
     6421 ggacaccgga ttataaatat tcggctgtag tacctgtagt tcagtataac aaatcatcta
     6481 ttgacggagt cccttatggt aaagcacata cattggcttt gaaaaagaat atccagttac
     6541 caacaaacac agaaaaagct tatcttagaa ctactatttc cggatgggga catgctaagc
     6601 catatgatgc gggaagcaga ggttgtgcag aatggtgctt cagaacacac actatagcaa
     6661 taaataattc gaatactttc cagcatcagc tgggtgcttt aggatgttca gcaaacccta
     6721 tcaataatca gagtccggga aattggactc ccgacagagc cggttggtgc ccgggaatgg
     6781 cagttccaac acgtatagat gtactgaata attctttaat aggcagtact tttagttatg
     6841 aatataaatt ccagaactgg acaaataacg gaaccaatgg agatgctttt tatgcaattt
     6901 ccagttttgt gattgcaaaa agtaatacac ctattagtgc tccggtagtt acaaactgag
     6961 gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta
     7021 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa aggaggaact
     7081 atatccggat atcccgcaag aggcccggca gtaccggcat aaccaagcct atgcctacag
     7141 catccagggt gacggtgccg aggatgacga tgagcgcatt gttagatttc atacacggtg
     7201 cctgactgcg ttagcaattt aactgtgata aactaccgca ttaaagctta tcgatgataa
     7261 gctgtc
//
