LOCUS       sequence-444332-        6456 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  IPTG-inducible expression of C. elegans protein ALDO-1 with a His 
            tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6456)
  AUTHORS   Katherine Walstrom Lab
  TITLE     Walstrom Lab Plasmids 2024
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6456)
  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            "Unpublished"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: ALDO-1_KS
FEATURES             Location/Qualifiers
     source          1..6456
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     110..127
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    142..281
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(207..226)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     367..389
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(383..574)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(1383..2465)
                     /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     2427..2446
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2466..2543)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     2749..2768
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2852..2871
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2852..2870
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2871..2895
                     /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             2911..2931
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4039..4056
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4109..4127)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4123..4170
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4207..4662
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4294..4313)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4504..4525
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4688..4792
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4793..4861,4862..5653)
                     /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     complement(5011..5030)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5824..6412
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6313..6332
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
       61 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
      121 gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
      181 ttcacaccgc aatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
      241 tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac ccgccaacac
      301 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
      361 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc
      421 agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct gcctgttcat
      481 ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg ataaagcggg
      541 ccatgttaag ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc
      601 tgttcatggg ggtaatgata ccgatgaaac gagagaggat gctcacgata cgggttactg
      661 atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa acaactggcg gtatggatgc
      721 ggcgggacca gagaaaaatc actcagggtc aatgccagcg cttcgttaat acagatgtag
      781 gtgttccaca gggtagccag cagcatcctg cgatgcagat ccggaacata atggtgcagg
      841 gcgctgactt ccgcgtttcc agactttacg aaacacggaa accgaagacc attcatgttg
      901 ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg
      961 attcattctg ctaaccagta aggcaacccc gccagcctag ccgggtcctc aacgacagga
     1021 gcacgatcat gcgcacccgt ggggccgcca tgccggcgat aatggcctgc ttctcgccga
     1081 aacgtttggt ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag attccgaata
     1141 ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg ccgaaaatga
     1201 cccagagcgc tgccggcacc tgtcctacga gttgcatgat aaagaagaca gtcataagtg
     1261 cggcgacgat agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca
     1321 agggcatcgg tcgagatccc ggtgcctaat gagtgagcta acttacatta attgcgttgc
     1381 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     1441 aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg tggtttttct tttcaccagt
     1501 gagacgggca acagctgatt gcccttcacc gcctggccct gagagagttg cagcaagcgg
     1561 tccacgctgg tttgccccag caggcgaaaa tcctgtttga tggtggttaa cggcgggata
     1621 taacatgagc tgtcttcggt atcgtcgtat cccactaccg agatatccgc accaacgcgc
     1681 agcccggact cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc
     1741 atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa accggacatg
     1801 gcactccagt cgccttcccg ttccgctatc ggctgaattt gattgcgagt gagatattta
     1861 tgccagccag ccagacgcag acgcgccgag acagaactta atgggcccgc taacagcgcg
     1921 atttgctggt gacccaatgc gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg
     1981 gagaaaataa tactgttgat gggtgtctgg tcagagacat caagaaataa cgccggaaca
     2041 ttagtgcagg cagcttccac agcaatggca tcctggtcat ccagcggata gttaatgatc
     2101 agcccactga cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc ttcgacgccg
     2161 cttcgttcta ccatcgacac caccacgctg gcacccagtt gatcggcgcg agatttaatc
     2221 gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac gccaatcagc
     2281 aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc
     2341 atcgccgctt ccactttttc ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg
     2401 cgggaaacgg tctgataaga gacaccggca tactctgcga catcgtataa cgttactggt
     2461 ttcacattca ccaccctgaa ttgactctct tccgggcgct atcatgccat accgcgaaag
     2521 gttttgcgcc attcgatggt gtccgggatc tcgacgctct cccttatgcg actcctgcat
     2581 taggaagcag cccagtagta ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc
     2641 atgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc
     2701 gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc
     2761 gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc
     2821 gtagaggatc gagatctcga tcccgcgaaa ttaatacgac tcactatagg ggaattgtga
     2881 gcggataaca attcccctct agtaataatt ttgtttaact ttaagaagga ggtctagatg
     2941 gcttcttact cgcaattctt gaccaaggcc caggaggacg aactccgctc catcgccaac
     3001 gccatcgtca ccccaggaaa gggaattctc gccgccgatg agtccaccgg atccatggac
     3061 aagcgtctta acagcatcgg actcgagaac accgaggaga accgccgcaa gtatcgtcaa
     3121 cttctgttta ccgccggagc cgacctcaac aagtacatct cgggagtcat catgttccac
     3181 gagacattct accaaaagac cgacgacgga aagccattca ctgctcttct ccaggagcaa
     3241 ggaatcattc caggaatcaa ggttgacaag ggtgtcgtcc caatggccgg aaccatcgga
     3301 gagggaacca ctcagggatt ggatgacttg aatgctcgtt gcgctcaata caagaaagac
     3361 ggagcacagt tcgccaagtg gagatgcgtg cacaagatct catctacgac tccatcagtg
     3421 actgctctta aggaaattgc ctccaacttg gctcgctatg cttcaatttg ccaacaaaat
     3481 ggacttgtac caattgttga gccagagatc ctcccagacg gtgaacactg cttggctcgc
     3541 ggacagaaga tcaccgagac cgtgctctca tatgtgtacc acgctctcaa cgagcatcac
     3601 gtcttcctcg agggaaccct cctcaagcca aacatggtca cctccggaca atcattcacc
     3661 ggcgagaagc catcgaatgc cgatatcgga ctcgccaccg tcaccgccct ccaacgcgga
     3721 gtcccatccg ccgttccagg agtcgtcttc ctctccggtg gacaatccga agaggatgcc
     3781 acattgaact tgaacgccat caatcaagtg agcggaaaga agccatgggc cctcacattc
     3841 tcctatggac gtgctcttca agcctcctgt ctcgccaagt gggccggaaa ggacgagaac
     3901 atcgcagccg cccaagaagt cctgctccat cgtgctcagg tcaactccct tgcctctgtc
     3961 ggaaaataca ccggggatgc ttcagctgac gcagccgctt cccaatccct cttcgtcgcc
     4021 aatcattctt atctcgagca ccaccaccac caccactgag atccggctgc taacaaagcc
     4081 cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg
     4141 gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc cggattggcg
     4201 aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     4261 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     4321 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     4381 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     4441 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     4501 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     4561 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     4621 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg
     4681 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     4741 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     4801 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     4861 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     4921 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     4981 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     5041 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     5101 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     5161 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     5221 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     5281 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgcagc
     5341 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     5401 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     5461 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     5521 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     5581 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     5641 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     5701 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     5761 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     5821 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     5881 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     5941 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     6001 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     6061 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     6121 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     6181 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     6241 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     6301 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     6361 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     6421 caacgcggcc tttttacggt tcctggcctt ttgctg
//
