LOCUS       sequence-197668-        6719 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses aldehyde dehydrogenase from Escherichia coli, codon 
            optimized for expression in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6719)
  TITLE     Siegel Lab ALDH Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6719)
  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            "Unpublished"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: ecALDH
FEATURES             Location/Qualifiers
     source          1..6719
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     893..912
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(932..1009)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1215..1234
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1322..1341
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1322..1340
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1341..1365
                     /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             1380..1402
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2901..2918
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2971..2989)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2985..3032
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3069..3524
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3156..3175)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3366..3387
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3616..4431)
                     /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(3616..4431)
                     /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     4339..4358
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4423..4442
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4553..5141
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5042..5061
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5295..5312
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5327..5466
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5392..5411)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5552..5574
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5568..5759)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6568..6719,1..931))
                     /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"
ORIGIN
        1 ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg cgaaaatcct
       61 gtttgatggt ggttaacggc gggatataac atgagctgtc ttcggtatcg tcgtatccca
      121 ctaccgagat gtccgcacca acgcgcagcc cggactcggt aatggcgcgc attgcgccca
      181 gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca ttcagcattt
      241 gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc gctatcggct
      301 gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc gccgagacag
      361 aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc agatgctcca
      421 cgcccagtcg cgtaccgtct tcatgggaga aaataatact gttgatgggt gtctggtcag
      481 agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca atggcatcct
      541 ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga agattgtgca
      601 ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc acgctggcac
      661 ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca
      721 gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt tgtgccacgc
      781 ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc gttttcgcag
      841 aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca ccggcatact
      901 ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga ctctcttccg
      961 ggcgctatca tgccataccg cgaaaggttt tgcgccattc gatggtgtcc gggatctcga
     1021 cgctctccct tatgcgactc ctgcattagg aagcagccca gtagtaggtt gaggccgttg
     1081 agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag tcccccggcc
     1141 acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc
     1201 cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc tgtggcgccg
     1261 gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga tcgatctcga tcccgcgaaa
     1321 ttaatacgac tcactatagg ggaattgtga gcggataaca attcccctct agaaataatt
     1381 ttgtttaact ttaagaagga gatatacata tgaacttcca ccatctggca tattggcagg
     1441 ataaagcact gagcctggca attgaaaatc gcctgtttat taacggtgaa tataccgcag
     1501 cagcagaaaa cgaaaccttt gaaaccgttg atccggttac ccaggcaccg ctggcaaaaa
     1561 ttgcacgtgg taaaagcgtt gatattgatc gtgcaatgag cgcagcacgt ggtgtttttg
     1621 aacgtggtga ttggagcctg agcagtccgg caaaacgtaa agcagttctg aataaactgg
     1681 ccgatctgat ggaagcacat gcagaagaac tggcactgct ggaaaccctg gataccggca
     1741 aaccgattcg tcatagcctg cgtgatgata ttccgggtgc agcccgtgca attcgttggt
     1801 atgcagaagc aattgataaa gtctatggtg aagttgcaac caccagtagc catgaactgg
     1861 caatgattgt tcgtgaacct gttggtgtta ttgcagcaat tgttccgtgg aattttccac
     1921 tgctgctgac ctgttggaaa ctgggtccgg cactggcagc aggtaatagc gttattctga
     1981 aaccgagcga aaaaagtccg ctgagcgcaa ttcgtctggc aggtctggca aaagaagcag
     2041 gtctgccgga cggtgttctg aatgttgtta ccggctttgg tcatgaagcc ggtcaggccc
     2101 tgagccgtca taacgatatt gatgcaattg cctttaccgg cagcacccgt acaggtaaac
     2161 agctgctgaa agatgccggt gatagcaata tgaaacgtgt ttggctggaa gccggtggta
     2221 aatcagcaaa tattgttttt gcagattgtc cggatctgca acaggcagca agcgcaaccg
     2281 cagccggtat tttctataat cagggtcagg tttgtattgc aggcacacgc ctgctgctgg
     2341 aagaaagcat tgcagatgaa tttctggccc tgctgaaaca gcaggcacag aattggcagc
     2401 ctggtcatcc gctggaccct gcaaccacaa tgggcaccct gattgattgt gcacatgccg
     2461 atagcgttca tagctttatt cgtgaaggtg aaagcaaagg tcagctgctg ctggatggtc
     2521 gtaatgccgg tctggcagcc gcaattggtc cgaccatttt tgttgatgtt gatccgaatg
     2581 caagcctgag tcgtgaagaa atttttggtc cggttctggt tgttacccgt tttaccagcg
     2641 aagaacaggc cctgcaactg gcaaacgata gccagtatgg tctgggtgca gcagtttgga
     2701 cccgtgatct gagccgtgca catcgtatga gccgtcgtct gaaagcaggt agcgtttttg
     2761 tgaataacta taacgatggt gatatgaccg ttccgtttgg tggttataaa cagagcggta
     2821 atggtcgtga taaaagcctg catgcactgg aaaaattcac cgaactgaaa accatttgga
     2881 ttagcctgga agcactcgag caccaccacc accaccactg agatccggct gctaacaaag
     2941 cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg
     3001 gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata tccggattgg
     3061 cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     3121 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     3181 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     3241 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     3301 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     3361 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     3421 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     3481 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg
     3541 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     3601 ctcatgaatt aattcttaga aaaactcatc gagcatcaaa tgaaactgca atttattcat
     3661 atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc
     3721 accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc
     3781 aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc
     3841 accatgagtg acgactgaat ccggtgagaa tggcaaaagt ttatgcattt ctttccagac
     3901 ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt
     3961 attcattcgt gattgcgcct gagcgagacg aaatacgcga tcgctgttaa aaggacaatt
     4021 acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc
     4081 acctgaatca ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt
     4141 gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa
     4201 ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt
     4261 gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc
     4321 acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt
     4381 ggaatttaat cgcggcctag agcaagacgt ttcccgttga atatggctca taacacccct
     4441 tgtattactg tttatgtaag cagacagttt tattgttcat gaccaaaatc ccttaacgtg
     4501 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     4561 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     4621 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     4681 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     4741 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     4801 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     4861 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     4921 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     4981 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     5041 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     5101 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     5161 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     5221 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     5281 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt
     5341 ttctccttac gcatctgtgc ggtatttcac accgcaatgg tgcactctca gtacaatctg
     5401 ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg
     5461 gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     5521 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     5581 ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc
     5641 gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt
     5701 aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact
     5761 gatgcctccg tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag
     5821 aggatgctca cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgat
     5881 ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc
     5941 cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg
     6001 cagatccgga acataatggt gcagggcgct gacttccgcg tttccagact ttacgaaaca
     6061 cggaaaccga agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg
     6121 cttcacgttc gctcgcgtat cggtgattca ttctgctaac cagtaaggca accccgccag
     6181 cctagccggg tcctcaacga caggagcacg atcatgcgca cccgtggggc cgccatgccg
     6241 gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga
     6301 gcgagggcgt gcaagattcc gaataccgca agcgacaggc cgatcatcgt cgcgctccag
     6361 cgaaagcggt cctcgccgaa aatgacccag agcgctgccg gcacctgtcc tacgagttgc
     6421 atgataaaga agacagtcat aagtgcggcg acgatagtca tgccccgcgc ccaccggaag
     6481 gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg
     6541 agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     6601 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     6661 cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcct
//
