LOCUS       sequence-462632-        6774 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Plasmodium vivax 6-phosphogluconate dehydrogeanse under T7
            with HIS tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6774)
  AUTHORS   Beckham J
  TITLE     Josh Beckham Lab plasmids 2024
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6774)
  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            "Unpublished"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: Pv6PGDH-pNic-Bsa4
FEATURES             Location/Qualifiers
     source          1..6774
                     /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             1639..1656
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1709..1727)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1723..1770
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1807..2262
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1894..1913)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2104..2125
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2354..3169)
                     /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(2354..3169)
                     /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     3077..3096
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3161..3180
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3291..3879
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3780..3799
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4033..4050
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4065..4204
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4130..4149)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4290..4312
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4306..4497)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5306..6388)
                     /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     6350..6369
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6389..6466)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6672..6691
                     /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 atccatgacc ggcacttgtg atatcggtct
      181 cattggtctg gcggttatgg gtcagaacct gtccctgaac atcgcctcta acggttttac
      241 catcggtgtt tacaaccgta cctacgagcg tacggaagat accctgaaga aagcgaaaga
      301 gggtaacctg ccgatccaag gttatgaaac cctcgagcag ctgatcaaca acctcaagaa
      361 accgcgtaaa atcattctgc tcatcaaagc gggtccggcg gttgacgaaa ccattaagaa
      421 catcctgaag cactttgagg aaggcgacat catcatcgac ggtggtaacg aatggtacct
      481 caataccgaa cgtcgtatca ctctgtgcga agagcacaaa gtagaatacc tggcgatggg
      541 tgtttctggt ggcgaagcag gtgcgcgcta tggctgttct ttcatgccgg gtggctccaa
      601 atatgcgtat gacacgatca aagacatcct cgagaagtgc tctgcgaaag ttggcacgag
      661 cccttgcgta acttacatcg gtcctcgtag ctctggtaac tacgttaaaa tggttcacaa
      721 cggtatcgaa tatggcgata tgcaactcat ctccgagtct tacctgctga tgaaaaacat
      781 tctgaactac aataacgaga aactgagcga agtattcaaa aaatggaacg aaggtatcct
      841 caactcttat ctgatcgaaa tcacctataa gattctgggc aagaaggacg aactcacgga
      901 caaccatctc gttgacatga tcctggacat cgcgggtgct aagggtacgg gtaaatggac
      961 catgctggaa gcgatcgagc gtggtatccc gtgcccgacc atgtgtgcgg cgctggacgc
     1021 ccgtaatatc tccgcgtata aacagctgcg cgttaaggcg gactctcatt tttctaacga
     1081 catgaaggtt aacaaagttg aaggtgaaaa tctcattaac tttgaagacg acctcctgaa
     1141 tgcgctgtac tgctgcaaga tcatttctta cacccagggt ctgttcctgc tcaaacaggt
     1201 ttctgaagag atgaagtgga atctcaatct gggtgaaatc tctcgtattt ggcgtggtgg
     1261 ctgcatcatc cgcgctgtct tcctggatcg tattactaac gcgtacaaga agaatgaaaa
     1321 gctcgacctg ctctttctcg acgaggattt cgcgcaggaa atgaagaaca aactcccgag
     1381 cctgcgtaaa gttgttcaag ttgcgaccaa atcttctatc ccgatcccgg cgttcagcgc
     1441 gtctctggcc tactttcaga tggtgacctc tcaaaacctc ccactgaacc tggtgcaggc
     1501 gcaacgtgac tacttcggct ctcacacgta taagcgtgtt gaccgtgacg gtgacttcca
     1561 tactatctgg gaataacagt aaaggtggat acggatccga attcgagctc cgtcgacaag
     1621 cttgcggccg cactcgagca ccaccaccac caccactgag atccggctgc taacaaagcc
     1681 cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg
     1741 gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc cggattggcg
     1801 aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     1861 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     1921 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     1981 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     2041 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     2101 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     2161 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     2221 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg
     2281 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     2341 catgaattaa ttcttagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat
     2401 caggattatc aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac
     2461 cgaggcagtt ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa
     2521 catcaataca acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac
     2581 catgagtgac gactgaatcc ggtgagaatg gcaaaagttt atgcatttct ttccagactt
     2641 gttcaacagg ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat
     2701 tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa ggacaattac
     2761 aaacaggaat cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac
     2821 ctgaatcagg atattcttct aatacctgga atgctgtttt cccggggatc gcagtggtga
     2881 gtaaccatgc atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt
     2941 ccgtcagcca gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc
     3001 catgtttcag aaacaactct ggcgcatcgg gcttcccata caatcgatag attgtcgcac
     3061 ctgattgccc gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg
     3121 aatttaatcg cggcctagag caagacgttt cccgttgaat atggctcata acaccccttg
     3181 tattactgtt tatgtaagca gacagtttta ttgttcatga ccaaaatccc ttaacgtgag
     3241 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     3301 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     3361 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     3421 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     3481 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     3541 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     3601 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     3661 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     3721 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     3781 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     3841 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     3901 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     3961 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     4021 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt
     4081 ctccttacgc atctgtgcgg tatttcacac cgcaatggtg cactctcagt acaatctgct
     4141 ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc
     4201 tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     4261 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     4321 gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga
     4381 ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa
     4441 tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga
     4501 tgcctccgtg taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag
     4561 gatgctcacg atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg
     4621 taaacaactg gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca
     4681 gcgcttcgtt aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca
     4741 gatccggaac ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg
     4801 gaaaccgaag accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct
     4861 tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc
     4921 tagccgggtc ctcaacgaca ggagcacgat catgcgcacc cgtggggccg ccatgccggc
     4981 gataatggcc tgcttctcgc cgaaacgttt ggtggcggga ccagtgacga aggcttgagc
     5041 gagggcgtgc aagattccga ataccgcaag cgacaggccg atcatcgtcg cgctccagcg
     5101 aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc acctgtccta cgagttgcat
     5161 gataaagaag acagtcataa gtgcggcgac gatagtcatg ccccgcgccc accggaagga
     5221 gctgactggg ttgaaggctc tcaagggcat cggtcgagat cccggtgcct aatgagtgag
     5281 ctaacttaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     5341 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca
     5401 gggtggtttt tcttttcacc agtgagacgg gcaacagctg attgcccttc accgcctggc
     5461 cctgagagag ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt
     5521 tgatggtggt taacggcggg atataacatg agctgtcttc ggtatcgtcg tatcccacta
     5581 ccgagatatc cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg
     5641 ccatctgatc gttggcaacc agcatcgcag tgggaacgat gccctcattc agcatttgca
     5701 tggtttgttg aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa
     5761 tttgattgcg agtgagatat ttatgccagc cagccagacg cagacgcgcc gagacagaac
     5821 ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc
     5881 ccagtcgcgt accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga
     5941 catcaagaaa taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt
     6001 catccagcgg atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg
     6061 ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca
     6121 gttgatcggc gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac
     6181 tggaggtggc aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt
     6241 tgggaatgta attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa
     6301 cgtggctggc ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg
     6361 cgacatcgta taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc
     6421 gctatcatgc cataccgcga aaggttttgc gccattcgat ggtgtccggg atctcgacgc
     6481 tctcccttat gcgactcctg cattaggaag cagcccagta gtaggttgag gccgttgagc
     6541 accgccgccg caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg
     6601 gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga
     6661 tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg
     6721 atgccggcca cgatgcgtcc ggcgtagagg atcgagatct cgatcccgcg aaat
//
