LOCUS       sequence-374851-        6268 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The construct was used to express recombinant ALDH1A3 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 6268)
  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 6268)
  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-ALDH1A3
FEATURES             Location/Qualifiers
     source          1..6268
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     510..529
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     613..632
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        613..631
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             663..685
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             705..722
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    732..756
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2329..2353)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2406..2424)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2420..2467
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(2833..2838,2839..2855,2856..2861))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(2888..2906)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2974..3078
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3079..3147,3148..3939)
                     /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     complement(3297..3316)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4110..4698
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4599..4618
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4852..4869
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4884..5023
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4949..4968)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5109..5131
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5125..5316)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 agacgtagcc cagcgcgtcg gccgccatgc cggcgataat ggcctgcttc tcgccgaaac
       61 gtttggtggc gggaccagtg acgaaggctt gagcgagggc gtgcaagatt ccgaataccg
      121 caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg gtcctcgccg aaaatgaccc
      181 agagcgctgc cggcacctgt cctacgagtt gcatgataaa gaagacagtc ataagtgcgg
      241 cgacgatagt catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg
      301 gcatcggtcg atcgacgctc tcccttatgc gactcctgca ttaggaagca gcccagtagt
      361 aggttgaggc cgttgagcac cgccgccgca aggaatggtg catgcaagga gatggcgccc
      421 aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc
      481 ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc
      541 gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggat cgagatctcg
      601 atcccgcgaa attaatacga ctcactatag ggagaccaca acggtttccc tctagaaata
      661 attttgttta actttaagaa ggagatatac atatgtcgta ctaccatcac catcaccatc
      721 acctcgaatc aacaagtttg tacaaaaaag caggctccgc ggccgccccc ttcaccgcca
      781 ccgctaacgg ggccgtggaa aacgggcagc cggacaggaa gccgccggcc ctgccgcgcc
      841 ccatccgcaa cctggaggtc aagttcacca agatatttat caacaatgaa tggcacgaat
      901 ccaagagtgg gaaaaagttt gctacatgta acccttcaac tcgggagcaa atatgtgaag
      961 tggaagaagg agataagccc gacgtggaca aggctgtgga ggctgcacag gttgccttcc
     1021 agaggggctc gccatggcgc cggctggatg ccctgagtcg tgggcggctg ctgcaccagc
     1081 tggctgacct ggtggagagg gaccgcgcca ccttggccgc cctggagacg atggatacag
     1141 ggaagccatt tcttcatgct tttttcatcg acctggaggg ctgtattaga accctcagat
     1201 actttgcagg gtgggcagac aaaatccagg gcaagaccat ccccacagat gacaacgtcg
     1261 tgtgcttcac caggcatgag cccattggtg tctgtggggc catcactcca tggaacttcc
     1321 ccctgctgat gctggtgtgg aagctggcac ccgccctctg ctgtgggaac accatggtcc
     1381 tgaagcctgc ggagcagaca cctctcaccg ccctttatct cggctctctg atcaaagagg
     1441 ccgggttccc tccaggagtg gtgaacattg tgccaggatt cgggcccaca gtgggagcag
     1501 caatttcttc tcaccctcag atcaacaaga tcgccttcac cggctccaca gaggttggaa
     1561 aactggttaa agaagctgcg tcccggagca atctgaagcg ggtgacgctg gagctggggg
     1621 ggaagaaccc ctgcatcgtg tgtgcggacg ctgacttgga cttggcagtg gagtgtgccc
     1681 atcagggagt gttcttcaac caaggccagt gttgcacggc agcctccagg gtgttcgtgg
     1741 aggagcaggt ctactctgag tttgtcaggc ggagcgtgga gtatgccaag aaacggcccg
     1801 tgggagaccc cttcgatgtc aaaacagaac aggggcctca gattgatcaa aagcagttcg
     1861 acaaaatctt agagctgatc gagagtggga agaaggaagg ggccaagctg gaatgcgggg
     1921 gctcagccat ggaagacaag gggctcttca tcaaacccac tgtcttctca gaagtcacag
     1981 acaacatgcg gattgccaaa gaggagattt tcgggccagt gcaaccaata ctgaagttca
     2041 aaagtatcga agaagtgata aaaagagcga atagcaccga ctatggactc acagcagccg
     2101 tgttcacaaa aaatctcgac aaagccctga agttggcttc tgccttagag tctggaacgg
     2161 tctggatcaa ctgctacaac gccctctatg cacaggctcc atttggtggc tttaaaatgt
     2221 caggaaatgg cagagaacta ggtgaatacg ctttggccga atacacagaa gtgaaaactg
     2281 tcaccatcaa acttggcgac aagaacccct gaaagggtgg gcgcgccgac ccagctttct
     2341 tgtacaaagt ggttgattcg aggctgctaa caaagcccga aaggaagctg agttggctgc
     2401 tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg
     2461 ttttttgctg aaaggaggaa ctatatccgg atatccacag gacgggtgtg gtcgccatga
     2521 tcgcgtagtc gatagtggct ccaagtagcg aagcgagcag gactgggcgg cggccaaagc
     2581 ggtcggacag tgctccgaga acgggtgcgc atagaaattg catcaacgca tatagcgcta
     2641 gcagcacgcc atagtgactg gcgatgctgt cggaatggac gatatcccgc aagaggcccg
     2701 gcagtaccgg cataaccaag cctatgccta cagcatccag ggtgacggtg ccgaggatga
     2761 cgatgagcgc attgttagat ttcatacacg gtgcctgact gcgttagcaa tttaactgtg
     2821 ataaactacc gcattaaagc ttatcgatga taagctgtca aacatgagaa ttcttgaaga
     2881 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     2941 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     3001 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     3061 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     3121 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     3181 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     3241 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     3301 tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac
     3361 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     3421 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     3481 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     3541 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     3601 gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact attaactggc
     3661 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     3721 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     3781 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     3841 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     3901 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     3961 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     4021 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     4081 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     4141 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     4201 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     4261 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     4321 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     4381 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     4441 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     4501 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     4561 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     4621 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     4681 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     4741 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     4801 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     4861 gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     4921 atttcacacc gcaatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     4981 agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac
     5041 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     5101 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     5161 gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc
     5221 atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg
     5281 ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt
     5341 tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac
     5401 tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat
     5461 gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt
     5521 aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca
     5581 gggcgctgac ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga ccattcatgt
     5641 tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg
     5701 tgattcattc tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag
     5761 gagcacgatc atgcgcaccc gtggccagga cccaacgctg cccgagatgc gccgcgtgcg
     5821 gctgctggag atggcggacg cgatggatat gttctgccaa gggttggttt gcgcattcac
     5881 agttctccgc aagaattgat tggctccaat tcttggagtg gtgaatccgt tagcgaggtg
     5941 ccgccggctt ccattcaggt cgaggtggcc cggctccatg caccgcgacg caacgcgggg
     6001 aggcagacaa ggtatagggc ggcgcctaca atccatgcca acccgttcca tgtgctcgcc
     6061 gaggcggcat aaatcgccgt gacgatcagc ggtccaatga tcgaagttag gctggtaaga
     6121 gccgcgagcg atccttgaag ctgtccctga tggtcgtcat ctacctgcct ggacagcatg
     6181 gcctgcaacg cgggcatccc gatgccgccg gaagcgagaa gaatcataat ggggaaggcc
     6241 atccagcctc gcgtcgcgaa cgccagca
//
