LOCUS       sequence-374862-        6235 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The construct was used to express recombinant ALDH1A1 in E.Coli. The
            purified protein was used for activity assay..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6235)
  AUTHORS   Li J, Garavaglia S, Ye Z, Moretti A, Belyaeva OV, Beiser A, Ibrahim 
            M, Wilk A, McClellan S, Klyuyeva AV, Goggans KR, Kedishvili NY, 
            Salter EA, Wierzbicki A, Migaud ME, Mullett SJ, Yates NA, Camacho 
            CJ, Rizzi M, Sobol RW
  TITLE     A specific inhibitor of ALDH1A3 regulates retinoic acid biosynthesis
            in glioma stem cells.
  JOURNAL   Commun Biol. 2021 Dec 21;4(1):1420. doi: 10.1038/s42003-021-02949-7.
  PUBMED    34934174
REFERENCE   2  (bases 1 to 6235)
  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: "Commun 
            Biol."; date: "2021-12-21"; volume: "4(1):1420. doi"; pages: " 
            10.1038/s42003-021-02949-7"
COMMENT     Sequence Label: pDEST17-ALDH1A1
FEATURES             Location/Qualifiers
     source          1..6235
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(96..115)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      909..1497
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1398..1417
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1651..1668
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1683..1822
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(1748..1767)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1908..1930
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1924..2115)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     3577..3596
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3680..3699
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3680..3698
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             3730..3752
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3772..3789
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    3799..3823
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(5363..5387)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(5440..5458)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5454..5501
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(5867..5872,5873..5889,5890..5895))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(5922..5940)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6008..6112
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6113..6181,6182..6235,1..738)
                     /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"
ORIGIN
        1 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
       61 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
      121 gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
      181 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
      241 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
      301 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
      361 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
      421 gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
      481 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
      541 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
      601 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
      661 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
      721 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
      781 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
      841 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
      901 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
      961 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     1021 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     1081 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     1141 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     1201 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     1261 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     1321 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     1381 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     1441 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     1501 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     1561 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     1621 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     1681 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg caatggtgca
     1741 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct
     1801 acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg
     1861 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     1921 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc
     1981 agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag
     2041 tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt
     2101 ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat
     2161 accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt
     2221 actggaacgt tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat
     2281 cactcagggt caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca
     2341 gcagcatcct gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc
     2401 cagactttac gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt
     2461 tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt
     2521 aaggcaaccc cgccagccta gccgggtcct caacgacagg agcacgatca tgcgcacccg
     2581 tggccaggac ccaacgctgc ccgagatgcg ccgcgtgcgg ctgctggaga tggcggacgc
     2641 gatggatatg ttctgccaag ggttggtttg cgcattcaca gttctccgca agaattgatt
     2701 ggctccaatt cttggagtgg tgaatccgtt agcgaggtgc cgccggcttc cattcaggtc
     2761 gaggtggccc ggctccatgc accgcgacgc aacgcgggga ggcagacaag gtatagggcg
     2821 gcgcctacaa tccatgccaa cccgttccat gtgctcgccg aggcggcata aatcgccgtg
     2881 acgatcagcg gtccaatgat cgaagttagg ctggtaagag ccgcgagcga tccttgaagc
     2941 tgtccctgat ggtcgtcatc tacctgcctg gacagcatgg cctgcaacgc gggcatcccg
     3001 atgccgccgg aagcgagaag aatcataatg gggaaggcca tccagcctcg cgtcgcgaac
     3061 gccagcaaga cgtagcccag cgcgtcggcc gccatgccgg cgataatggc ctgcttctcg
     3121 ccgaaacgtt tggtggcggg accagtgacg aaggcttgag cgagggcgtg caagattccg
     3181 aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa
     3241 atgacccaga gcgctgccgg cacctgtcct acgagttgca tgataaagaa gacagtcata
     3301 agtgcggcga cgatagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct
     3361 ctcaagggca tcggtcgatc gacgctctcc cttatgcgac tcctgcatta ggaagcagcc
     3421 cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat
     3481 ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct
     3541 catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc
     3601 agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatcga
     3661 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
     3721 agaaataatt ttgtttaact ttaagaagga gatatacata tgtcgtacta ccatcaccat
     3781 caccatcacc tcgaatcaac aagtttgtac aaaaaagcag gctccgcggc cgcccccttc
     3841 acctcatcct caggcacgcc agacttacct gtcctactca ccgatttgaa gattcaatat
     3901 actaagatct tcataaacaa tgaatggcat gattcagtga gtggcaagaa atttcctgtc
     3961 tttaatcctg caactgagga ggagctctgc caggtagaag aaggagataa ggaggatgtt
     4021 gacaaggcag tgaaggccgc aagacaggct tttcagattg gatccccgtg gcgtactatg
     4081 gatgcttccg agagggggcg actattatac aagttggctg atttaatcga aagagatcgt
     4141 ctgctgctgg cgacaatgga gtcaatgaat ggtggaaaac tctattccaa tgcatatctg
     4201 aatgatttag caggctgcat caaaacattg cgctactgtg caggttgggc tgacaagatc
     4261 cagggccgta caataccaat tgatggaaat ttttttacat atacaagaca tgaacctatt
     4321 ggtgtatgtg gccaaatcat tccttggaat ttcccgttgg ttatgctcat ttggaagata
     4381 gggcctgcac tgagctgtgg aaacacagtg gttgtcaaac cagcagagca aactcctctc
     4441 actgctctcc acgtggcatc tttaataaaa gaggcagggt ttcctcctgg agtagtgaat
     4501 attgttcctg gttatgggcc tacagcaggg gcagccattt cttctcacat ggatatagac
     4561 aaagtagcct tcacaggatc aacagaggtt ggcaagttga tcaaagaagc tgccgggaaa
     4621 agcaatctga agagggtgac cctggagctt ggaggaaaga gcccttgcat tgtgttagct
     4681 gatgccgact tggacaatgc tgttgaattt gcacaccatg gggtattcta ccaccagggc
     4741 cagtgttgta tagccgcatc caggattttt gtggaagaat caatttatga tgagtttgtt
     4801 cgaaggagtg ttgagcgggc taagaagtat atccttggaa atcctctgac cccaggagtc
     4861 actcaaggcc ctcagattga caaggaacaa tatgataaaa tactcgacct cattgagagt
     4921 gggaagaaag aaggggccaa actggaatgt ggaggaggcc cgtgggggaa taaaggctac
     4981 tttgtccagc ccacagtgtt ctctaatgtt acagatgaga tgcgcattgc caaagaggag
     5041 atttttggac cagtgcagca aatcatgaag tttaaatctt tagatgacgt gatcaaaaga
     5101 gcaaacaata ctttctatgg cttatcagca ggagtgttta ccaaagacat tgataaagcc
     5161 ataacaatct cctctgctct gcaggcagga acagtgtggg tgaattgcta tggcgtggta
     5221 agtgcccagt gcccctttgg tggattcaag atgtctggaa atggaagaga actgggagag
     5281 tacggtttcc atgaatatac agaggtcaaa acagtcacag tgaaaatctc tcagaagaac
     5341 tcataaaagg gtgggcgcgc cgacccagct ttcttgtaca aagtggttga ttcgaggctg
     5401 ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat
     5461 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat
     5521 ccggatatcc acaggacggg tgtggtcgcc atgatcgcgt agtcgatagt ggctccaagt
     5581 agcgaagcga gcaggactgg gcggcggcca aagcggtcgg acagtgctcc gagaacgggt
     5641 gcgcatagaa attgcatcaa cgcatatagc gctagcagca cgccatagtg actggcgatg
     5701 ctgtcggaat ggacgatatc ccgcaagagg cccggcagta ccggcataac caagcctatg
     5761 cctacagcat ccagggtgac ggtgccgagg atgacgatga gcgcattgtt agatttcata
     5821 cacggtgcct gactgcgtta gcaatttaac tgtgataaac taccgcatta aagcttatcg
     5881 atgataagct gtcaaacatg agaattcttg aagacgaaag ggcctcgtga tacgcctatt
     5941 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     6001 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     6061 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     6121 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     6181 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacga
//
