LOCUS       sequence-444358-        6928 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  IPTG-inducible expression of C. elegans protein GSPD-1 with a His 
            tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6928)
  AUTHORS   Loges LN, Walstrom KM
  TITLE     Modeling human glucose-6-phosphate dehydrogenase mutations using C. 
            elegans GSPD-1.
  JOURNAL   MicroPubl Biol. 2021 Sep 10;2021:10.17912/micropub.biology.000451. 
            doi: 10.17912/micropub.biology.000451. eCollection 2021.
  PUBMED    34532700
REFERENCE   2  (bases 1 to 6928)
  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: "MicroPubl 
            Biol."; date: "2021-09-10"; volume: 
            "2021:10.17912/micropub.biology.000451. doi"; pages: " 
            10.17912/micropub.biology.000451. eCollection 2021"
COMMENT     Sequence Label: GSPD1_AJ
FEATURES             Location/Qualifiers
     source          1..6928
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(11..439)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(121..142)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     333..352
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(476..523)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     519..537
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(590..607)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             2187..2207
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    2223..2247
                     /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     complement(2247..2266)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2248..2266)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2350..2369)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        2575..2652
                     /gene="lacI"
                     /label=lacI promoter
     CDS             2653..3735
                     /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(2672..2691)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             4544..4735
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(4729..4751)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4837..4976
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4892..4911
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4991..5008)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5162..5750)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5242..5261)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5921..6712,6713..6781))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6544..6563
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6782..6886)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
       61 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
      121 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
      181 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
      241 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
      301 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
      361 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
      421 gcgccgctac agggcgcgtc ccattcgcca atccggatat agttcctcct ttcagcaaaa
      481 aacccctcaa gacccgttta gaggccccaa ggggttatgc tagttattgc tcagcggtgg
      541 cagcagccaa ctcagcttcc tttcgggctt tgttagcagc cggatctcag tggtggtggt
      601 ggtggtgctc gagtagcttt ggagcaaccc atttgtatgt tccagtgaag atgaaaccgt
      661 actttttcat caattcgtca ccttcagttg gaccacgact tccaaattta tattgaaccg
      721 gttgaacttt cttcttttta agctcttcga gaactggcgt caaaatacgc caagcgtatt
      781 caagctcatc agttcgaacg aaattaattt gagatcccat gaatacttcg agaaataggc
      841 gttcataagc atcaggaaga cggacttctt tgaatctatt attataagtc aaatcaagtt
      901 cagtttcttc aacaccgaat cccattccag gtttcttggt catcagtttc atgtaaactg
      961 cctcatttgg ttgcactctc atcactaatt cggatcgctt taattcacca gacggataga
     1021 tatctccact gacttctttg aattgtatac gaacttctgc tttcttctcg ttgagagctt
     1081 ttccacaacg caagaagaat ggaacaccct cccaacgttc attattgatg tgaacaacag
     1141 caagagcata cgttggagta gtcgaatcgg ctggaactga tttgtcatct ttgtatcctt
     1201 gtgatgcttc tgggtgatca aattcaggac tggcaatgta ttgtccaact actacatcct
     1261 ttaattctac aactttagca gctttcaaca ctttgacctt ttcatcacga atatcttcag
     1321 cattcaaact tgctggtttc tccatggcga ctaaagttag aatttgcatc aaatgattct
     1381 gcataacatc acggataatt ccagctgtat caaaatatcc agcacggcca ccagttccaa
     1441 aatcttcttt aaatgagatc ataactgatg caatgtgatc gcggttccaa gatggagcga
     1501 gaattcggtt tccaaatcgc ataaccatca aattctgaac catttctttt ccgagataat
     1561 gatcaatacg gtagatttga tcttctttaa aaagtttcgc caaatgagtt gaaagttcac
     1621 aagatgattt caaatcatgt ccaaatggtt tttctatgat aacacgtgtc catgaatcac
     1681 catgatccat acaattcttc tttaattccg tagaaaccac gttgaaaact gatggaggaa
     1741 gagcgaggta gtagagacga ttgacggctt gattattgct ttctttttgg aaatcatcaa
     1801 ttgaagattg aaggcgttgg aatccttcag aagtgtcgta ttggccctga acgtatgagc
     1861 acttcttgat gaagtcatca aaagcacatt tctcattttc tcgaacttta cagtttttct
     1921 caaaagattc acgaagcttg aaaactgtca gatcggaacg ggcatatcca ataaatttga
     1981 tgttgactgg caaaaggttg tcacggaaca accaccacag tgttgggtat attttctttt
     2041 ttgcgagatc tcccgatgct ccaaatatca cgaaaacgta cggagtctca aatttgagat
     2101 ctctttgcat tgattcacga agacactcga cgagatcctt cgaaatggga tcggacactg
     2161 aatgacgttt gcatgccatt ctagacctcc ttcttaaagt taaacaaaat tattactaga
     2221 ggggaattgt tatccgctca caattcccct atagtgagtc gtattaattt cgcgggatcg
     2281 agatctcgat cctctacgcc ggacgcatcg tggccggcat caccggcgcc acaggtgcgg
     2341 ttgctggcgc ctatatcgcc gacatcaccg atggggaaga tcgggctcgc cacttcgggc
     2401 tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc cgtggccggg ggactgttgg
     2461 gcgccatctc cttgcatgca ccattccttg cggcggcggt gctcaacggc ctcaacctac
     2521 tactgggctg cttcctaatg caggagtcgc ataagggaga gcgtcgagat cccggacacc
     2581 atcgaatggc gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga gagtcaattc
     2641 agggtggtga atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc cggtgtctct
     2701 tatcagaccg tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa
     2761 aaagtggaag cggcgatggc ggagctgaat tacattccca accgcgtggc acaacaactg
     2821 gcgggcaaac agtcgttgct gattggcgtt gccacctcca gtctggccct gcacgcgccg
     2881 tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac tgggtgccag cgtggtggtg
     2941 tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg
     3001 caacgcgtca gtgggctgat cattaactat ccgctggatg accaggatgc cattgctgtg
     3061 gaagctgcct gcactaatgt tccggcgtta tttcttgatg tctctgacca gacacccatc
     3121 aacagtatta ttttctccca tgaagacggt acgcgactgg gcgtggagca tctggtcgca
     3181 ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg
     3241 cgtctggctg gctggcataa atatctcact cgcaatcaaa ttcagccgat agcggaacgg
     3301 gaaggcgact ggagtgccat gtccggtttt caacaaacca tgcaaatgct gaatgagggc
     3361 atcgttccca ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc
     3421 attaccgagt ccgggctgcg cgttggtgcg gatatctcgg tagtgggata cgacgatacc
     3481 gaagacagct catgttatat cccgccgtta accaccatca aacaggattt tcgcctgctg
     3541 gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt gaagggcaat
     3601 cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc
     3661 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     3721 gaaagcgggc agtgagcgca acgcaattaa tgtaagttag ctcactcatt aggcaccggg
     3781 atctcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg
     3841 catgactatc gtcgccgcac ttatgactgt cttctttatc atgcaactcg taggacaggt
     3901 gccggcagcg ctctgggtca ttttcggcga ggaccgcttt cgctggagcg cgacgatgat
     3961 cggcctgtcg cttgcggtat tcggaatctt gcacgccctc gctcaagcct tcgtcactgg
     4021 tcccgccacc aaacgtttcg gcgagaagca ggccattatc gccggcatgg cggccccacg
     4081 ggtgcgcatg atcgtgctcc tgtcgttgag gacccggcta ggctggcggg gttgccttac
     4141 tggttagcag aatgaatcac cgatacgcga gcgaacgtga agcgactgct gctgcaaaac
     4201 gtctgcgacc tgagcaacaa catgaatggt cttcggtttc cgtgtttcgt aaagtctgga
     4261 aacgcggaag tcagcgccct gcaccattat gttccggatc tgcatcgcag gatgctgctg
     4321 gctaccctgt ggaacaccta catctgtatt aacgaagcgc tggcattgac cctgagtgat
     4381 ttttctctgg tcccgccgca tccataccgc cagttgttta ccctcacaac gttccagtaa
     4441 ccgggcatgt tcatcatcag taacccgtat cgtgagcatc ctctctcgtt tcatcggtat
     4501 cattaccccc atgaacagaa atccccctta cacggaggca tcagtgacca aacaggaaaa
     4561 aaccgccctt aacatggccc gctttatcag aagccagaca ttaacgcttc tggagaaact
     4621 caacgagctg gacgcggatg aacaggcaga catctgtgaa tcgcttcacg accacgctga
     4681 tgagctttac cgcagctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     4741 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     4801 tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag
     4861 cgatagcgga gtgtatactg gcttaactat gcggcatcag agcagattgt actgagagtg
     4921 caccattgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgct
     4981 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     5041 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     5101 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     5161 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     5221 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     5281 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     5341 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     5401 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     5461 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     5521 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     5581 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     5641 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     5701 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     5761 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     5821 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     5881 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     5941 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     6001 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     6061 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     6121 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     6181 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctgcag
     6241 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     6301 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     6361 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     6421 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     6481 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     6541 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     6601 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     6661 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     6721 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     6781 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     6841 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     6901 aagtgccacc taaattgtaa gcgttaat
//
