LOCUS       sequence-443144-        6733 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of human aldolase A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6733)
  AUTHORS   Fenton KD, Meneely KM, Wu T, Martin TA, Swint-Kruse L, Fenton AW, 
            Lamb AL
  TITLE     Substitutions at a rheostat position in human aldolase A cause a 
            shift in the conformational population.
  JOURNAL   Protein Sci. 2022 Feb;31(2):357-370. doi: 10.1002/pro.4222. Epub 
            2021 Nov 12.
  PUBMED    34734672
REFERENCE   2  (bases 1 to 6733)
  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"; doi: "10.1002/pro.4222"; 
            journalName: "Protein Sci"; date: "2022-02"; volume: "31"; issue: 
            "2"; pages: "357-370"
COMMENT     Sequence Label: pET11a-AldoA
FEATURES             Location/Qualifiers
     source          1..6733
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        186..263
                     /gene="lacI"
                     /label=lacI promoter
     CDS             264..1346
                     /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(283..302)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             2658..2849
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(2843..2865)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2951..3090
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3006..3025
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3105..3122)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3276..3864)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3356..3375)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4035..4826,4827..4895))
                     /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     4658..4677
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4896..5000)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5068..5086
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(5113..5118,5119..5135,5136..5141)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     terminator      complement(5316..5363)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     5359..5377
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     RBS             6530..6552
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    6567..6591
                     /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(6591..6610)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6592..6610)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6694..6713)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 ccacttcggg ctcatgagcg cttgtttcgg cgtgggtatg gtggcaggcc ccgtggccgg
       61 gggactgttg ggcgccatct ccttgcatgc accattcctt gcggcggcgg tgctcaacgg
      121 cctcaaccta ctactgggct gcttcctaat gcaggagtcg cataagggag agcgtcgaga
      181 tcccggacac catcgaatgg cgcaaaacct ttcgcggtat ggcatgatag cgcccggaag
      241 agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt atacgatgtc gcagagtatg
      301 ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca ggccagccac gtttctgcga
      361 aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg
      421 cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc agtctggccc
      481 tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca
      541 gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca
      601 atcttctcgc gcaacgcgtc agtgggctga tcattaacta tccgctggat gaccaggatg
      661 ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt atttcttgat gtctctgacc
      721 agacacccat caacagtatt attttctccc atgaagacgg tacgcgactg ggcgtggagc
      781 atctggtcgc attgggtcac cagcaaatcg cgctgttagc gggcccatta agttctgtct
      841 cggcgcgtct gcgtctggct ggctggcata aatatctcac tcgcaatcaa attcagccga
      901 tagcggaacg ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc
      961 tgaatgaggg catcgttccc actgcgatgc tggttgccaa cgatcagatg gcgctgggcg
     1021 caatgcgcgc cattaccgag tccgggctgc gcgttggtgc ggatatctcg gtagtgggat
     1081 acgacgatac cgaagacagc tcatgttata tcccgccgtt aaccaccatc aaacaggatt
     1141 ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca actctctcag ggccaggcgg
     1201 tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag aaaaaccacc ctggcgccca
     1261 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     1321 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtaagtta gctcactcat
     1381 taggcaccgg gatctcgacc gatgcccttg agagccttca acccagtcag ctccttccgg
     1441 tgggcgcggg gcatgactat cgtcgccgca cttatgactg tcttctttat catgcaactc
     1501 gtaggacagg tgccggcagc gctctgggtc attttcggcg aggaccgctt tcgctggagc
     1561 gcgacgatga tcggcctgtc gcttgcggta ttcggaatct tgcacgccct cgctcaagcc
     1621 ttcgtcactg gtcccgccac caaacgtttc ggcgagaagc aggccattat cgccggcatg
     1681 gcggccgacg cgctgggcta cgtcttgctg gcgttcgcga cgcgaggctg gatggccttc
     1741 cccattatga ttcttctcgc ttccggcggc atcgggatgc ccgcgttgca ggccatgctg
     1801 tccaggcagg tagatgacga ccatcaggga cagcttcaag gatcgctcgc ggctcttacc
     1861 agcctaactt cgatcattgg accgctgatc gtcacggcga tttatgccgc ctcggcgagc
     1921 acatggaacg ggttggcatg gattgtaggc gccgccctat accttgtctg cctccccgcg
     1981 ttgcgtcgcg gtgcatggag ccgggccacc tcgacctgaa tggaagccgg cggcacctcg
     2041 ctaacggatt caccactcca agaattggag ccaatcaatt cttgcggaga actgtgaatg
     2101 cgcaaaccaa cccttggcag aacatatcca tcgcgtccgc catctccagc agccgcacgc
     2161 ggcgcatctc gggcagcgtt gggtcctggc cacgggtgcg catgatcgtg ctcctgtcgt
     2221 tgaggacccg gctaggctgg cggggttgcc ttactggtta gcagaatgaa tcaccgatac
     2281 gcgagcgaac gtgaagcgac tgctgctgca aaacgtctgc gacctgagca acaacatgaa
     2341 tggtcttcgg tttccgtgtt tcgtaaagtc tggaaacgcg gaagtcagcg ccctgcacca
     2401 ttatgttccg gatctgcatc gcaggatgct gctggctacc ctgtggaaca cctacatctg
     2461 tattaacgaa gcgctggcat tgaccctgag tgatttttct ctggtcccgc cgcatccata
     2521 ccgccagttg tttaccctca caacgttcca gtaaccgggc atgttcatca tcagtaaccc
     2581 gtatcgtgag catcctctct cgtttcatcg gtatcattac ccccatgaac agaaatcccc
     2641 cttacacgga ggcatcagtg accaaacagg aaaaaaccgc ccttaacatg gcccgcttta
     2701 tcagaagcca gacattaacg cttctggaga aactcaacga gctggacgcg gatgaacagg
     2761 cagacatctg tgaatcgctt cacgaccacg ctgatgagct ttaccgcagc tgcctcgcgc
     2821 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     2881 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     2941 ggtgtcgggg cgcagccatg acccagtcac gtagcgatag cggagtgtat actggcttaa
     3001 ctatgcggca tcagagcaga ttgtactgag agtgcaccat tgcggtgtga aataccgcac
     3061 agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg cttcctcgct cactgactcg
     3121 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     3181 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     3241 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     3301 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     3361 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     3421 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     3481 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     3541 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     3601 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     3661 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     3721 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     3781 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     3841 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     3901 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     3961 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     4021 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     4081 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     4141 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     4201 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     4261 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     4321 aatagtttgc gcaacgttgt tgccattgct gcaggcatcg tggtgtcacg ctcgtcgttt
     4381 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     4441 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     4501 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     4561 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     4621 cggcgaccga gttgctcttg cccggcgtca acacgggata ataccgcgcc acatagcaga
     4681 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     4741 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     4801 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     4861 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     4921 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     4981 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     5041 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtcttcaa
     5101 gaattctcat gtttgacagc ttatcatcga taagctttaa tgcggtagtt tatcacagtt
     5161 aaattgctaa cgcagtcagg caccgtgtat gaaatctaac aatgcgctca tcgtcatcct
     5221 cggcaccgtc accctggatg ctgtaggcat aggcttggtt atgccggtac tgccgggcct
     5281 cttgcgggat atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta
     5341 gaggccccaa ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc
     5401 tttcgggctt tgttagcagc cggatcctta ataggcgtgg ttagagacga agagggactc
     5461 gctggcagca gccccagcct gaccgctcgg agtgtacttt ccttgacagg caaggctgtt
     5521 ggccagggct cgcttgacat actcctcctg cgcagccttc aggttctcct tcttcccgcc
     5581 ccaggccttc agggcagagg cctgcagggc tcggccgtag gagaaggtca gggcccaggg
     5641 cttcagcagg gggcacttgt taatggcatt gaggttgatg gacgcctcct cctcactctg
     5701 gcctccagac aggaaggtga tcccagtgac agcggggggc actgtgcggc gcagcgctgt
     5761 gacggtcgcc atggcaatct cctcatgaga aaacttctga gtgcaagcat ggcctggggt
     5821 gaccatgttg ggcttcagca aggtgccttc caggtagatg tggtggtcac tcagagcctt
     5881 gtagacagca gccagcacct tctcggtcac atactggcag cgcttcaagt catggtcccc
     5941 atcagggagg atctcaggct ccacgatggg cacaatgcca ttctgctggc agatactggc
     6001 ataacgggcc agaacattgg cattttccat gatggcgagg gctgaggggg tgtgttcccc
     6061 aatcttcagc acacaacgcc acttggcgaa gtcagctccg tccttcttgt actgggcaca
     6121 gcgctcagac agcccatcca acccttgggt ggtagtctcg ccatttgtcc ctgccagggg
     6181 gaccacgccc ttgtctacct tgatgcccac aacaccgccc ttggatttga taacttgggg
     6241 gaagggacgc ccatcatccg ccttctggta gagtgtctca tggaagagga tgacaccccc
     6301 aatgcagggg ttcacgcggt cgtcagctgt cagcagcagc tggcggtaga agcgccggtt
     6361 ctcctcggtg ttctcggtgc caatggactg cagccgcttg gcaatgctcc cagtggactc
     6421 atctgcagcc aggatgccct tgccaggtgc cacgatgcgg tgagcgatgt cagacagctc
     6481 cttcttctgc tccggggtca gtgctggata ttggtagggc atatgtatat ctccttctta
     6541 aagttaaaca aaattatttc tagaggggaa ttgttatccg ctcacaattc ccctatagtg
     6601 agtcgtatta atttcgcggg atcgagatct cgatcctcta cgccggacgc atcgtggccg
     6661 gcatcaccgg cgccacaggt gcggttgctg gcgcctatat cgccgacatc accgatgggg
     6721 aagatcgggc tcg
//
