LOCUS       sequence-214870-        7818 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression and purification of ALDOA_Glycolytic.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7818)
  AUTHORS   Teyra J, Kelil A, Jain S, Helmy M, Jajodia R, Hooda Y, Gu J, D'Cruz 
            AA, Nicholson SE, Min J, Sudol M, Kim PM, Bader GD, Sidhu SS
  TITLE     Large-scale survey and database of high affinity ligands for peptide
            recognition modules.
  JOURNAL   Mol Syst Biol. 2020 Dec;16(12):e9310. doi: 10.15252/msb.20199310.
  PUBMED    33438817
REFERENCE   2  (bases 1 to 7818)
  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 Syst 
            Biol."; date: "2020-12"; volume: "16(12):e9310. doi"; pages: " 
            10.15252/msb.20199310"
COMMENT     Sequence Label: pHH0103_ALDOA_Glycolytic
FEATURES             Location/Qualifiers
     source          1..7818
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..6,7..22,23..29)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    37..53
                     /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)."
     primer_bind     42..64
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             138..155
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             171..824
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     782..804
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             831..854
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             867..884
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     terminator      2013..2060
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      complement(3653..4108)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3790..3811)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4002..4021
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(4150..4172)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4258..4398
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4313..4332
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4413..4430)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4584..5171)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5342..6133,6134..6202))
                     /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     5965..5984
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6203..6307)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6375..6393
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6416..6493
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             6494..7576
                     /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     complement(6513..6532)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ttgacaatta atcatcggct cgtataatgt gtggaattgt gagcggataa caatttcaca
       61 caggaaacag ccagtccgtt taggtgtttt cacgagcact tcaccaacaa ggaccataga
      121 ttatgaaaat cgaagaacac catcaccatc accattccag cggtaagctt atgtccccta
      181 tactaggtta ttggaaaatt aagggccttg tgcaacccac tcgacttctt ttggaatatc
      241 ttgaagaaaa atatgaagag catttgtatg agcgcgatga aggtgataaa tggcgaaaca
      301 aaaagtttga attgggtttg gagtttccca atcttcctta ttatattgat ggtgatgtta
      361 aattaacaca gtctatggcc atcatacgtt atatagctga caagcacaac atgttgggtg
      421 gttgtccaaa agagcgtgca gagatttcaa tgcttgaagg agcggttttg gatattagat
      481 acggtgtttc gagaattgca tatagtaaag actttgaaac tctcaaagtt gattttctta
      541 gcaagctacc tgaaatgctg aaaatgttcg aagatcgttt atgtcataaa acatatttaa
      601 atggtgatca tgtaacccat cctgacttca tgttgtatga cgctcttgat gttgttttat
      661 acatggaccc aatgtgcctg gatgcgttcc caaaattagt ttgttttaaa aaacgtattg
      721 aagctatccc acaaattgat aagtacttga aatccagcaa gtatatagca tggcctttgc
      781 agggctggca agccacgttt ggtggtggcg accatcctcc aaaatcggat ctggaagttc
      841 tgttccaggg gcccctgtcc agcggtctgg ttccgcgtgg ttccggtacc gcggcccagc
      901 cggccatgcc gtatcagtac ccggcgctga cgccggaaca gaaaaaagaa ctgagtgaca
      961 ttgcccatcg tatcgttgca ccgggcaaag gtattctggc ggccgatgaa agtaccggct
     1021 ccattgccaa acgcctgcag tccatcggta cggaaaacac cgaagaaaat cgtcgctttt
     1081 atcgtcaact gctgctgacg gccgatgacc gcgtcaaccc gtgcattggc ggtgtgatcc
     1141 tgtttcacga aaccctgtac cagaaagcag atgacggccg tccgttcccg caagttatta
     1201 aatctaaggg cggtgtggtt ggcatcaaag tcgacaaggg tgtcgtgccg ctggcaggca
     1261 cgaatggtga aaccacgacc cagggcctgg atggtctgag cgaacgttgc gcgcaatata
     1321 aaaaggacgg cgccgatttc gcaaaatggc gctgtgtgct gaagattggt gaacataccc
     1381 cgtcagccct ggcaatcatg gaaaacgcga atgttctggc tcgttatgcg tcgatttgcc
     1441 agcaaaacgg cattgttccg atcgtcgaac cggaaatcct gccggatggt gaccacgatc
     1501 tgaaacgctg tcagtacgtg acggaaaaag ttctggcagc tgtctataag gcgctgagcg
     1561 atcatcacat ttacctggaa ggcaccctgc tgaaaccgaa tatggtcacg ccgggtcatg
     1621 cctgtaccca gaaattttct cacgaagaaa ttgctatggc aacggtgacc gcactgcgtc
     1681 gcaccgtgcc gccggcagtt acgggtatca ccttcctgtc aggcggtcaa tcggaagaag
     1741 aagcgagtat taacctgaat gcgatcaaca aatgcccgct gctgaagccg tgggctctga
     1801 cctttagcta tggtcgtgcg ctgcaggcct ctgcactgaa agcctggggc ggtaaaaagg
     1861 aaaacctgaa agcggcccag gaagaatatg ttaagcgcgc tctggcgaat agtctggcat
     1921 gtcagggtaa atacaccccg tccggtcaag ctggtgcagc tgcgagcgaa tctctgttcg
     1981 tgagcaatca tgcgtacgcg gccgcataat aaaccgatac aattaaaggc tccttttgga
     2041 gccttttttt tggagatttt caacgtgaaa aaattattat tcgcaattcc tttagttgtt
     2101 cctttctatt ctcactccgc tgaaactgtt gaaagttgtt tagcaaaacc ccatacagaa
     2161 aattcattta ctaacgtctg gaaagacgac aaaactttag atcgttacgc taactatgag
     2221 ggctgtctgt ggaatgctac aggcgttgta gtttgtactg gtgacgaaac tcagtgtcta
     2281 gctagagtgg cggtggctct ggttccggtg attttgatta tgaaaagatg gcaaacgcta
     2341 ataagggggc tatgaccgaa aatgccgatg aaaacgcgct acagtctgac gctaaaggca
     2401 aacttgattc tgtcgctact gattacggtg ctgctatcga tggtttcatt ggtgacgttt
     2461 ccggccttgc taatggtaat ggtgctactg gtgattttgc tggctctaat tcccaaatgg
     2521 ctcaagtcgg tgacggtgat aattcacctt taatgaataa tttccgtcaa tatttacctt
     2581 ccctccctca atcggttgaa tgtcgccctt ttgtctttgg cgctggtaaa ccatatgaat
     2641 tttctattga ttgtgacaaa ataaacttat tccgtggtgt ctttgcgttt cttttatatg
     2701 ttgccacctt tatgtatgta ttttctacgt ttgctaacat actgcgtaat aaggagtctt
     2761 aatcatgcca gttcttttgg ctagcgccgc cctatacctt gtctgcctcc ccgcgttgcg
     2821 tcgcggtgca tggagccggg ccacctcgac ctgaatggaa gccggcggca cctcgctaac
     2881 ggattcacca ctccaagaat tggagccaat caattcttgc ggagaactgt gaatgcgcaa
     2941 accaaccctt ggcagaacat atccatcgcg tccgccatct ccagcagccg cacgcggcgc
     3001 atctcgggca gcgttgggtc ctggccacgg gtgcgcatga tcgtgctcct gtcgttgagg
     3061 acccggctag gctggcgggg ttgccttact ggttagcaga atgaatcacc gatacgcgag
     3121 cgaacgtgaa gcgactgctg ctgcaaaacg tctgcgacct gagcaacaac atgaatggtc
     3181 ttcggtttcc gtgtttcgta aagtctggaa acgcggaagt cagcgccctg caccattatg
     3241 ttccggatct gcatcgcagg atgctgctgg ctaccctgtg gaacacctac atctgtatta
     3301 acgaagcgct ggcattgacc ctgagtgatt tttctctggt cccgccgcat ccataccgcc
     3361 agttgtttac cctcacaacg ttccagtaac cgggcatgtt catcatcagt aacccgtatc
     3421 gtgagcatcc tctctcgttt catcggtatc attaccccca tgaacagaaa tcccccttac
     3481 acggaggcat cagtgaccaa acaggaaaaa accgccctta acatggcccg ctttatcaga
     3541 agccagacat taacgcttct ggagaaactc aacgagctgg acgcggatga acaggcagac
     3601 atctgtgaat cgcttcacga ccacgctgat gagctttacc gcaggatccg cgaaattgta
     3661 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     3721 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     3781 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     3841 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     3901 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     3961 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     4021 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     4081 gcgcttaatg cgccgctaca gggcgcgtcg cggatcctgc ctcgcgcgtt tcggtgatga
     4141 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     4201 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc
     4261 agccatgacc cagtcacgta gcgatagcgg agtgtatact ggcttaacta tgcggcatca
     4321 gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg
     4381 agaaaatacc gcatcaggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     4441 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     4501 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4561 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     4621 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccagggtttc
     4681 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4741 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4801 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4861 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4921 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     4981 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     5041 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     5101 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     5161 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     5221 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     5281 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     5341 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     5401 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     5461 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     5521 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     5581 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5641 acgttgttgc cattgctgca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5701 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     5761 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     5821 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     5881 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     5941 gctcttgccc ggcgtcaaca cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     6001 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     6061 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     6121 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     6181 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     6241 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     6301 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     6361 cattaaccta taaaaatagg cgtatcacga ggccctttcg tcttcaagaa ttcccgacac
     6421 catcgaatgg tgcaaaacct ttcgcggtat ggcatgatag cgcccggaag agagtcaatt
     6481 cagggtggtg aatgtgaaac cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc
     6541 ttatcagacc gtttcccgcg tggtgaacca ggccagccac gtttctgcga aaacgcggga
     6601 aaaagtggaa gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg cacaacaact
     6661 ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc agtctggccc tgcacgcgcc
     6721 gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt
     6781 gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc
     6841 gcaacgcgtc agtgggctga tcattaacta tccgctggat gaccaggatg ccattgctgt
     6901 ggaagctgcc tgcactaatg ttccggcgtt atttcttgat gtctctgacc agacacccat
     6961 caacagtatt attttctccc atgaagacgg tacgcgactg ggcgtggagc atctggtcgc
     7021 attgggtcac cagcaaatcg cgctgttagc gggcccatta agttctgtct cggcgcgtct
     7081 gcgtctggct ggctggcata aatatctcac tcgcaatcaa attcagccga tagcggaacg
     7141 ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg
     7201 catcgttccc actgcgatgc tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc
     7261 cattaccgag tccgggctgc gcgttggtgc ggatatctcg gtagtgggat acgacgatac
     7321 cgaagacagc tcatgttata tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct
     7381 ggggcaaacc agcgtggacc gcttgctgca actctctcag ggccaggcgg tgaagggcaa
     7441 tcagctgttg cccgtctcac tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac
     7501 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     7561 ggaaagcggg cagtgagcgc aacgcaatta atgtaagtta gctcactcat taggcacaat
     7621 tctcatgttt gacagcttat catcgactgc acggtgcacc aatgcttctg gcgtcaggca
     7681 gccatcggaa gctgtggtat ggctgtgcag gtcgtaaatc actgcataat tcgtgtcgct
     7741 caaggcgcac tcccgttctg gataatgttt tttgcgccga catcataacg gttctggcaa
     7801 atattctgaa atgagctg
//
