LOCUS       sequence-398434-        3313 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains Magnet flanked by linkers and BsaI sites insert for 
            transposon insertion libraries..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3313)
  AUTHORS   Tague N, Andreani V, Fan Y, Timp W, Dunlop MJ
  TITLE     Comprehensive Screening of a Light-Inducible Split Cre Recombinase 
            with Domain Insertion Profiling.
  JOURNAL   ACS Synth Biol. 2023 Oct 3. doi: 10.1021/acssynbio.3c00328.
  PUBMED    37788288
REFERENCE   2  (bases 1 to 3313)
  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: "ACS Synth 
            Biol. 2023 Oct 3. doi: 10.1021/acssynbio.3c00328."
COMMENT     Sequence Label: pATT-BsaI-Pmag-NmagHigh1
FEATURES             Location/Qualifiers
     source          1..3313
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     265..284
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     518..535
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     634..653
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        634..652
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             684..706
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             762..1211
                     /codon_start=1
                     /product="""positive Magnet"" variant of an N-terminally 
                     truncated Vivid (VVD) blue light photoreceptor from 
                     Neurospora crassa (Kawano et al., 2015)"
                     /label=pMag
                     /note="rapidly heterodimerizes with nMagHigh1 upon blue 
                     light irradiation to form a complex with slow switch-off 
                     kinetics"
                     /translation="HTLYAPGGYDIMGYLRQIRNRPNPQVELGPVDTSCALILCDLKQK
                     DTPIVYASEAFLYMTGYSNAEVLGRNCRFLQSPDGMVKPKSTRKYVDSNTINTMRKAID
                     RNAEVQVEVVNFKKNGQRFVNFLTMIPVRDETGEYRYSMGFQCETE"
     RBS             1224..1235
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     primer_bind     complement(1786..1804)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1800..1847
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1953..2057
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2058..2126,2127..2918)
                     /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(2276..2295)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(3089..3313,1..364)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
       61 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
      121 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
      181 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
      241 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
      301 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
      361 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
      421 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
      481 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
      541 ggaagagcgc catcagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg
      601 gcgtagagga tcgagatctc gatcccgcga aattaatacg actcactata gggacaccac
      661 aacggtttcc ctctagaaat aattttgttt aactttaaga aggagatata ccatcgtctc
      721 tgtccggtct cgcatcgggt tctggaggct caagcggatc acacactctt tacgcccctg
      781 gaggatacga cattatggga tatttgcggc agattaggaa ccgcccaaac cctcaggtcg
      841 aactggggcc tgtggacacg tcatgtgccc tgatcctgtg cgatctgaag caaaaggaca
      901 ctccgatcgt ctacgcctcg gaagccttct tgtatatgac cggatacagc aatgcagagg
      961 tgctcggcag gaactgcaga ttcctgcagt cccccgacgg gatggtgaaa ccaaagtcga
     1021 ctcgcaaata tgtggactcg aacacgatca acaccatgcg gaaggccatc gaccggaacg
     1081 ccgaggtcca ggtggaggtg gtcaacttta agaagaacgg ccagcggttc gtgaactttc
     1141 tgaccatgat tccggtccgg gatgaaaccg gagagtacag atactccatg ggattccagt
     1201 gcgaaacaga ataagaattc attaaagagg agaaaggtac catgcataca ctttacgctc
     1261 ctgggggcta cgacatcatg ggctatttgg atcagattgg caatcgcccg aatccacagg
     1321 ttgaattagg gccagtcgat acgtcgtgcg cactgatttt gtgtgattta aagcaaaagg
     1381 ataccccaat tgtttacgcg agtgaggcgt ttctgtatat gacgggctac tcaaatgcgg
     1441 aggtacttgg ccgcaactgt cgcttcttac aatcgccgga cggcatggta aagcctaagt
     1501 caactcgtaa atacgttgac tccaacacta tcaatacaat tcgcaaagcg atcgatcgca
     1561 acgcagaggt ccaggtggag gttgttaact ttaagaagaa tgggcaacgc ttcgtgaatt
     1621 ttcttacgat tattccggtt cgtgacgaaa ccggcgaata tcgttactct atggggttcc
     1681 agtgtgaaac cgaaggtggc ggaggtagcg cgtcgagacc agcgagagac ggttaagatc
     1741 cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac
     1801 tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaaggaggaa
     1861 ctatatccgg atatccacag gacgggtgtg gtcgccatga tcgcgtagtc aaggtttctt
     1921 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     1981 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     2041 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     2101 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     2161 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     2221 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     2281 gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact
     2341 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     2401 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     2461 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     2521 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     2581 agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     2641 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     2701 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     2761 ccggtgagcg tggctctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     2821 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     2881 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     2941 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     3001 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     3061 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     3121 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     3181 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     3241 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     3301 ctctgctaat cct
//
