LOCUS       sequence-233164-        5826 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector containing gene for EhPTP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5826)
  TITLE     EhPTP-pNIC-Bsa4
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5826)
  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            "Unpublished"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: EhPTP-pNIC-Bsa4
FEATURES             Location/Qualifiers
     source          1..5826
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             92..109
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             134..154
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             691..708
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(761..779)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      775..822
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      859..1314
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(946..965)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1156..1177
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1406..2221)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1406..2221)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2129..2148
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2213..2232
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2343..2931
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2832..2851
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3085..3102
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3117..3256
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3182..3201)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3342..3364
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3358..3549)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4358..5440)
                     /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     5402..5421
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5441..5518)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5724..5743
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat gcaccatcat catcatcatt cttctggtgt
      121 agatctgggt accgagaacc tgtacttcca atccatgaaa ctgctgttcg tttgcctggg
      181 taacatctgc cgttctccgg ctgcggaagc ggttatgaaa aaagttatcc agaaccacca
      241 cctgaccgaa aaatacatct gtgactctgc gggtacctgc tcttaccacg aaggtcagca
      301 ggcggactct cgtatgcgta aagttggtaa atctcgtggt taccaggttg actctatctc
      361 tcgtccggtt gtttcttctg acttcaagaa ctttgactac atcttcgcga tggacaacga
      421 caactactac gaactcctgg accgttgccc ggaacagtac aaacagaaaa tcttcaaaat
      481 ggtagacttc tgcaccacca tcaaaaccac cgaagttccg gacccgtact acggtggtga
      541 aaaaggtttc caccgtgtta tcgacatcct ggaggacgcg tgcgaaaacc tgatcatcaa
      601 actggaagaa ggtaaactga tcaactaaca gtaaaggtgg atacggatcc gaattcgagc
      661 tccgtcgaca agcttgcggc cgcactcgag caccaccacc accaccactg agatccggct
      721 gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca
      781 taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata
      841 tccggattgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
      901 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
      961 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     1021 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     1081 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     1141 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     1201 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     1261 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg
     1321 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     1381 tatgtatccg ctcatgaatt aattcttaga aaaactcatc gagcatcaaa tgaaactgca
     1441 atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag
     1501 gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc
     1561 cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa
     1621 gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagt ttatgcattt
     1681 ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa
     1741 ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcga tcgctgttaa
     1801 aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa
     1861 caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt ttcccgggga
     1921 tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa
     1981 gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa
     2041 cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat
     2101 agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag
     2161 catccatgtt ggaatttaat cgcggcctag agcaagacgt ttcccgttga atatggctca
     2221 taacacccct tgtattactg tttatgtaag cagacagttt tattgttcat gaccaaaatc
     2281 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     2341 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     2401 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     2461 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     2521 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     2581 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     2641 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     2701 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     2761 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     2821 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     2881 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     2941 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     3001 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     3061 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg
     3121 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcaatgg tgcactctca
     3181 gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga
     3241 ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     3301 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     3361 gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg
     3421 gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc
     3481 cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg
     3541 tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa tgataccgat
     3601 gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc ggttactgga
     3661 acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa aaatcactca
     3721 gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta gccagcagca
     3781 tcctgcgatg cagatccgga acataatggt gcagggcgct gacttccgcg tttccagact
     3841 ttacgaaaca cggaaaccga agaccattca tgttgttgct caggtcgcag acgttttgca
     3901 gcagcagtcg cttcacgttc gctcgcgtat cggtgattca ttctgctaac cagtaaggca
     3961 accccgccag cctagccggg tcctcaacga caggagcacg atcatgcgca cccgtggggc
     4021 cgccatgccg gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg gaccagtgac
     4081 gaaggcttga gcgagggcgt gcaagattcc gaataccgca agcgacaggc cgatcatcgt
     4141 cgcgctccag cgaaagcggt cctcgccgaa aatgacccag agcgctgccg gcacctgtcc
     4201 tacgagttgc atgataaaga agacagtcat aagtgcggcg acgatagtca tgccccgcgc
     4261 ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc
     4321 ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     4381 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     4441 tattgggcgc cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct
     4501 tcaccgcctg gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc
     4561 gaaaatcctg tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt
     4621 cgtatcccac taccgagata tccgcaccaa cgcgcagccc ggactcggta atggcgcgca
     4681 ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat
     4741 tcagcatttg catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg
     4801 ctatcggctg aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg
     4861 ccgagacaga acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca
     4921 gatgctccac gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg
     4981 tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa
     5041 tggcatcctg gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa
     5101 gattgtgcac cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca
     5161 cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt
     5221 gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt
     5281 gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg
     5341 ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac
     5401 cggcatactc tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac
     5461 tctcttccgg gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggtgtccg
     5521 ggatctcgac gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg
     5581 aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc gcccaacagt
     5641 cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag
     5701 tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct
     5761 gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg
     5821 cgaaat
//
