LOCUS       sequence-486843-        4233 bp    DNA     circular SYN 22-SEP-2025
DEFINITION  Gateway middle entry clone containing a light inducible Cre 
            Recombinase (LiCre). .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4233)
  AUTHORS   Gillespie W, Zhang Y, Ruiz OE, Cerda J, Ortiz-Guzman J, Turner WD, 
            Largoza G, Sherman M, Mosser LE, Fujimoto E, Chien CB, Kwan KM, 
            Arenkiel BR, Devine WP, Wythe JD
  TITLE     Multisite Assembly of Gateway Induced Clones (MAGIC): a flexible 
            cloning toolbox with diverse applications in vertebrate model 
            systems.
  JOURNAL   bioRxiv [Preprint]. 2024 Jul 13:2024.07.13.603267. doi: 
            10.1101/2024.07.13.603267.
  PUBMED    39026881
REFERENCE   2  (bases 1 to 4233)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 22, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            [Preprint]. 2024 Jul 13:2024.07.13.603267. doi: 
            10.1101/2024.07.13.603267."
COMMENT     Sequence Label: pME-LiCre (JDW 1119)
FEATURES             Location/Qualifiers
     source          1..4233
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     670..686
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        696..714
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      750..759
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             762..782
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             786..806
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             819..836
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2258..2274)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2258..2274)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2311..2331)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2311..2329)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    complement(2358..2457)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(2475..2493)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2498..2514)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2498..2514)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2616..2635)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2627..3436
                     /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="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2694..2713)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3583..4171
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4072..4091
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggcttg taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat
      721 tgggtaccgg gccccccctc gaggtcgacg ccaccatggg tccaaaaaag aagagaaagg
      781 tagatccaaa aaagaagaga aaggtaatga ggggcagcca ccaccaccac caccacggca
      841 gcctggccac caccctggag aggatcgaga agaacttcgt gatcaccgac cccaggctgc
      901 ccgacaaccc catcatcttc gccagcgaca gcttcctgca gctgaccgag tacagcaggg
      961 aggagatcct gggcaggaac tgcaggttcc tgcagggccc cgagaccgac agggccaccg
     1021 tgaggaagat cagggacgcc atcgacaacc agaccgaggt gaccgtgcag ctgatcaact
     1081 acaccaagag cggcaagaag ttctggaacg tgttccacct gcagcccatg agggactaca
     1141 agggcgacgt gcagtacttc atcggcgtgc agctggacgg caccgagagg ctgcacggcg
     1201 ccgccgagag ggaggccgtg tgcctgatca agaagaccgc cttccagatc gacagcgacg
     1261 aggtgaggaa gaacctgatg gacatgttca gggacaggca ggccttcagc gagcacacct
     1321 ggaagatgct gctgagcgtg tgcaggagct gggccgcctg gtgcaagctg aacaacagga
     1381 agtggttccc cgccgagccc gaggacgtga gggactacct gctgtacctg caggccaggg
     1441 gcctggccgt gaagaccatc cagcagcacc tgggccagct gaacatgctg cacaggagga
     1501 gcggcctgcc caggcccagc gacagcaacg ccgtgagcct ggtgatgagg aggatcagga
     1561 aggagaacgt ggacgccggc gagagggcca agcaggccct ggccttcgag aggaccgact
     1621 tcgaccaggt gaggagcctg atggagaaca gcgacaggtg ccaggacatc aggaacctgg
     1681 ccttcctggg catcgcctac aacaccctgc tgaggatcgc cgagatcgcc aggatcaggg
     1741 tgaaggacat cagcaggacc gacggcggca ggatgctgat ccacatcggc aggaccaaga
     1801 ccctggtgag caccgccggc gtggagaagg ccctgagcct gggcgtgacc aagctggtgg
     1861 agaggtggat cagcgtgagc ggcgtggccg acgaccccaa caactacctg ttctgcaggg
     1921 tgaggaagaa cggcgtggcc gcccccagcg ccaccagcca gctgagcacc agggccctgg
     1981 agggcatctt cgaggccacc cacaggctga tctacggcgc caaggacgac agcggccaga
     2041 ggtacctggc ctggagcggc cacagcgcca gggtgggcgc cgccagggac atggccaggg
     2101 ccggcgtgag catccccgag atcatgcagg ccggcggctg gaccaacgtg aacatcgtga
     2161 tgaactacat caggaacctg gacagcgaga ccggcgccat ggtgaggctg ctggccgccg
     2221 gcgactaagc ttgatatcga attcctgcag cccgggggat ccactagttc tagagcggcc
     2281 gccaccgcgg tggagctcca gcttttgttc cctttagtga gggttaattg cgcgcttggc
     2341 gtaatcatgg tcatagcacc cagctttctt gtacaaagtt ggcattataa gaaagcattg
     2401 cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca
     2461 tccagctgat atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct
     2521 ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg
     2581 aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca
     2641 acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa
     2701 atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc
     2761 cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga
     2821 tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt
     2881 tatccgtact cctgatgatg catggttact caccactgcg atccccggaa aaacagcatt
     2941 ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt
     3001 cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt
     3061 tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga
     3121 tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc
     3181 attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga
     3241 cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca
     3301 ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct
     3361 ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct
     3421 cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct
     3481 gacttgacgg gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt
     3541 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     3601 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     3661 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     3721 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     3781 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     3841 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     3901 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     3961 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     4021 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     4081 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     4141 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     4201 cctggccttt tgctggcctt ttgctcacat gtt
//
