LOCUS       sequence-403108-        4404 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression plasmid with shRNAmiR targeting BACH1, with a BFP 
            reporter on the backbone. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4404)
  AUTHORS   Wan Y, Cohen J, Szenk M, Farquhar KS, Coraci D, Krzyszton R, Azukas 
            J, Van Nest N, Smashnov A, Chern YJ, De Martino D, Nguyen LC, Bien 
            H, Bravo-Cordero JJ, Chan CH, Rosner MR, Balazsi G
  TITLE     Nonmonotone invasion landscape by noise-aware control of metastasis 
            activator levels.
  JOURNAL   Nat Chem Biol. 2023 Jul;19(7):887-899. doi: 
            10.1038/s41589-023-01344-z. Epub 2023 May 25.
  PUBMED    37231268
REFERENCE   2  (bases 1 to 4404)
  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: 
            "10.1038/s41589-023-01344-z"; journalName: "Nat Chem Biol"; date: 
            "2023-07"; volume: "19"; issue: "7"; pages: "887-899"
COMMENT     Sequence Label: pUC-hU6-shBACH1_BFP
FEATURES             Location/Qualifiers
     source          1..4404
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     ncRNA           109..123
                     /label=miR-30a loop
                     /note="loop from the the 71-nt precursor of the human 
                     miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     promoter        226..1404
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          457..1395
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1352..1372
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     complement(2147..2164)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2153..2377
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        2404..2508
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2509..2577,2578..3369)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2727..2746)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3540..4128
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4029..4048
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(4191..4404,1..35)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     4191..4211
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     4362..4381
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 tggctttata tatcttgtgg aaaggacgcg acttcttaac ccaacagaag gctcgagaag
       61 gtatattgct gttgacagtg agcgcaacga tgattatgtt tcagaatagt gaagccacag
      121 atgtattctg aaacataatc atcgttttgc ctactgcctc ggacttcaag gggctagaat
      181 tcgttttttt ttgtatcaag tactagaact ccccagtggg aattcggctc cggtgcccgt
      241 cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat
      301 tgaaccggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg
      361 ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac
      421 gttctttttc gcaacgggtt tgccgccaga acacaggtaa gtgccgtgtg tggttcccgc
      481 gggcctggcc tctttacggg ttatggccct tgcgtgcctt gaattacttc cacctggctg
      541 cagtacgtga ttcttgatcc cgagcttcgg gttggaagtg ggtgggagag ttcgaggcct
      601 tgcgcttaag gagccccttc gcctcgtgct tgagttgagg cctggcctgg gcgctggggc
      661 cgccgcgtgc gaatctggtg gcaccttcgc gcctgtctcg ctgctttcga taagtctcta
      721 gccatttaaa atttttgatg acctgctgcg acgctttttt tctggcaaga tagtcttgta
      781 aatgcgggcc aagatctgca cactggtatt tcggtttttg gggccgcggg cggcgacggg
      841 gcccgtgcgt cccagcgcac atgttcggcg aggcggggcc tgcgagcgcg gccaccgaga
      901 atcggacggg ggtagtctca agctggccgg cctgctctgg tgcctggcct cgcgccgccg
      961 tgtatcgccc cgccctgggc ggcaaggctg gcccggtcgg caccagttgc gtgagcggaa
     1021 agatggccgc ttcccggccc tgctgcaggg agctcaaaat gaaggacgcg gcgctcggga
     1081 gagcgggcgg gtgagtcacc cacacaaagg aaaagggcct ttccgtcctc agccgtcgct
     1141 tcatgtgact ccacggagta ccgggcgccg tccaggcacc tcgattagtt ctcgagcttt
     1201 tggagtacgt cgtctttagg ttggggggag gggttttatg cgatggagtt tccccacact
     1261 gagtgggtgg agactgaagt taggccagct tggcacttga tgtaattctc cttggaattt
     1321 gccctttttg agtttggatc ttggttcatt ctcaagcctc agacagtggt tcaaagtttt
     1381 tttcttccat ttcaggtgtc gtgacgtacg gccaccatga gcatgtccga actgattaaa
     1441 gagaacatgc acatgaaact ttatatggaa ggtacggtcg acaaccacca tttcaaatgt
     1501 acctccgaag gagagggcaa accctacgag ggcacccaaa cgatgagaat taaagtagta
     1561 gaaggtgggc cgctgccctt tgcctttgac attctggcaa cttcttttct ctacgggagt
     1621 aaaactttta tcaaccatac gcaaggcatt ccagacttct ttaagcagtc atttcccgag
     1681 gggtttactt gggagcgggt cacaacatac gaggacggcg gcgtgctgac cgctactcag
     1741 gacaccagcc tgcaagacgg gtgtttgatc tacaatgtga agattcgagg cgtaaacttt
     1801 acttcaaacg gcccagttat gcagaaaaaa accctcggct gggaggcttt tacagagact
     1861 ctttacccag ccgacggtgg tttggaaggc cgaaacgaca tggcattgaa attggttggc
     1921 gggagccacc ttatagcgaa tataaagacg acctatcggt ctaaaaagcc cgccaaaaat
     1981 ttgaagatgc ctggtgtata ttacgtcgac tataggcttg agagaataaa agaagctaac
     2041 aacgaaacct atgtggagca acatgaggtt gcagttgcac gctactgcga tctgccgagc
     2101 aaattggggc acaagctgaa ttgatcagct agagctcgct aatcagcctc gactgtgcct
     2161 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     2221 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     2281 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagat
     2341 aatagcaggc atgctgggga tgcggtgggc tctatggtct agatttaggg taggggaaat
     2401 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     2461 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     2521 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     2581 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     2641 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     2701 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     2761 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     2821 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     2881 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     2941 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     3001 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     3061 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     3121 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     3181 gctggctggt ttattgctga taaatctgga gccggtgagc gtggaagccg cggtatcatt
     3241 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     3301 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     3361 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     3421 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     3481 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     3541 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     3601 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     3661 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     3721 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     3781 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     3841 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     3901 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     3961 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     4021 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     4081 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     4141 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcagctagc gagggcctat
     4201 ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag ataattggaa
     4261 ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga aagtaataat
     4321 ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat atgcttaccg
     4381 taacttgaaa gtatttcgat ttct
//
