LOCUS       sequence-268073-        6060 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Middle entry vector (Tol2 kit) with bi-cistronic CreLite system 
            components, PhyB-Delta-CreC and PIF6CreN, linked by P2A..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6060)
  AUTHORS   Yen ST, Trimmer KA, Aboul-Fettouh N, Mullen RD, Culver JC, Dickinson
            ME, Behringer RR, Eisenhoffer GT
  TITLE     CreLite: An optogenetically controlled Cre/loxP system using red 
            light.
  JOURNAL   Dev Dyn. 2020 Nov;249(11):1394-1403. doi: 10.1002/dvdy.232. Epub 
            2020 Aug 31.
  PUBMED    32745301
REFERENCE   2  (bases 1 to 6060)
  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: "10.1002/dvdy.232"; 
            journalName: "Dev Dyn"; date: "2020-11"; volume: "249"; issue: "11";
            pages: "1394-1403"
COMMENT     Sequence Label: pME-CreLite
FEATURES             Location/Qualifiers
     source          1..6060
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..100
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             112..132
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2962..3018
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             3022..3042
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    complement(3617..3716)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3733..3752)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3734..3752)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3757..3773)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3757..3773)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3875..3894)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3886..4695
                     /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(3953..3972)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4842..5430
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5331..5350
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5584..5601
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(5760..5787)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(5879..5965)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     6014..6036
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6028..6045
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6029..6045
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacaaat tgatgagcaa
       61 tgctttttta taatgccaac tttgtacaaa aaagcaggct tagcaaacat gcccaagaag
      121 aagaggaagg tggtttccgg agtcgggggt agtggcggtg gccgtggcgg tggccgtggc
      181 ggagaagaag aaccgtcgtc aagtcacact cctaataacc gaagaggagg agaacaagct
      241 caatcgtcgg gaacgaaatc tctcagacca agaagcaaca ctgaatcaat gagcaaagca
      301 attcaacagt acaccgtcga cgcaagactc cacgccgttt tcgaacaatc cggcgaatca
      361 gggaaatcat tcgactactc acaatcactc aaaacgacga cgtacggttc ctctgtacct
      421 gagcaacaga tcacagctta tctctctcga atccagcgag gtggttacat tcagcctttc
      481 ggatgtatga tcgccgtcga tgaatccagt ttccggatca tcggttacag tgaaaacgcc
      541 agagaaatgt tagggattat gcctcaatct gttcctactc ttgagaaacc tgagattcta
      601 gctatgggaa ctgatgtgag atctttgttc acttcttcga gctcgattct actcgagcgt
      661 gctttcgttg ctcgagagat taccttgtta aatccggttt ggatccattc caagaatact
      721 ggtaaaccgt tttacgccat tcttcatagg attgatgttg gtgttgttat tgatttagag
      781 ccagctagaa ctgaagatcc tgcgctttct attgctggtg ctgttcaatc gcagaaactc
      841 gcggttcgtg cgatttctca gttacaggct cttcctggtg gagatattaa gcttttgtgt
      901 gacactgtcg tggaaagtgt gagggacttg actggttatg atcgtgttat ggtttataag
      961 tttcatgaag atgagcatgg agaagttgta gctgagagta aacgagacga tttagagcct
     1021 tatattggac tgcattatcc tgctactgat attcctcaag cgtcaaggtt cttgtttaag
     1081 cagaaccgtg tccgaatgat agtagattgc aatgccacac ctgttcttgt ggtccaggac
     1141 gataggctaa ctcagtctat gtgcttggtt ggttctactc ttagggctcc tcatggttgt
     1201 cactctcagt atatggctaa catgggatct attgcgtctt tagcaatggc ggttataatc
     1261 aatggaaatg aagatgatgg gagcaatgta gctagtggaa gaagctcgat gaggctttgg
     1321 ggtttggttg tttgccatca cacttcttct cgctgcatac cgtttccgct aaggtatgct
     1381 tgtgagtttt tgatgcaggc tttcggttta cagttaaaca tggaattgca gttagctttg
     1441 caaatgtcag agaaacgcgt tttgagaacg cagacactgt tatgtgatat gcttctgcgt
     1501 gactcgcctg ctggaattgt tacacagagt cccagtatca tggacttagt gaaatgtgac
     1561 ggtgcagcat ttctttacca cgggaagtat tacccgttgg gtgttgctcc tagtgaagtt
     1621 cagataaaag atgttgtgga gtggttgctt gcgaatcatg cggattcaac cggattaagc
     1681 actgatagtt taggcgatgc ggggtatccc ggtgcagctg cgttagggga tgctgtgtgc
     1741 ggtatggcag ttgcatatat cacaaaaaga gactttcttt tttggtttcg atctcacact
     1801 gcgaaagaaa tcaaatgggg aggcgctaag catcatccgg aggataaaga tgatgggcaa
     1861 cgaatgcatc ctcgttcgtc ctttcaggct tttcttgaag ttgttaagag ccggagtcag
     1921 ccatgggaaa ctgcggaaat ggatgcgatt cactcgctcc agcttattct gagagactct
     1981 tttaaagaat ctggtggttc tgccggtggc tccggttctg gctccagcgg tggcagctct
     2041 ggtgcgtccg gcacgggtac tgcgggtggc actggcagcg gttccggtac tggctctggc
     2101 aaccggaaat ggtttcccgc agaacctgaa gatgttcgcg attatcttct atatcttcag
     2161 gcgcgcggtc tggcagtaaa aactatccag caacatttgg gccagctaaa catgcttcat
     2221 cgtcggtccg ggctgccacg accaagtgac agcaatgctg tttcactggt tatgcggcgg
     2281 atccgaaaag aaaacgttga tgccggtgaa cgtgcaaaac aggctctagc gttcgaacgc
     2341 actgatttcg accaggttcg ttcactcatg gaaaatagcg atcgctgcca ggatatacgt
     2401 aatctggcat ttctggggat tgcttataac accctgttac gtatagccga aattgccagg
     2461 atcagggtta aagatatctc acgtactgac ggtgggagaa tgttaatcca tattggcaga
     2521 acgaaaacgc tggttagcac cgcaggtgta gagaaggcac ttagcctggg ggtaactaaa
     2581 ctggtcgagc gatggatttc cgtctctggt gtagctgatg atccgaataa ctacctgttt
     2641 tgccgggtca gaaaaaatgg tgttgccgcg ccatctgcca ccagccagct atcaactcgc
     2701 gccctggaag ggatttttga agcaactcat cgattgattt acggcgctaa ggatgactct
     2761 ggtcagagat acctggcctg gtctggacac agtgcccgtg tcggagccgc gcgagatatg
     2821 gcccgcgctg gagtttcaat accggagatc atgcaagctg gtggctggac caatgtaaat
     2881 attgtcatga actatatccg taacctggat agtgaaacag gggcaatggt gcgcctgctg
     2941 gaagatggcg atggaagcgg agctactaac ttcagcctgc tgaagcaggc tggagacgtg
     3001 gaggagaacc ctggacctat gcccaagaag aagaggaagg tgatgttctt accaaccgat
     3061 tattgttgca ggttaagcga tcaagagtat atggagcttg tgtttgagaa tggccagatt
     3121 cttgcaaagg gccaaagatc caacgtttct ctgcataatc aacgtaccaa atcgatcatg
     3181 gatttgtatg aggcagagta taacgaggat ttcatgaaga gtatcatcca tggtggtggt
     3241 ggtgccatca caaatctcgg ggacacgcag gttgttccac aaagtcatgt tgctgctgcc
     3301 catgaaacaa acatgttgga aagcaataaa catgttgacg gtggttctgc cggtggctcc
     3361 ggttctggct ccagcggtgg cagctctggt gcgtccggca cgggtactgc gggtggcact
     3421 ggcagcggtt ccggtactgg ctctggcctg actgtgcacc aaaacctgcc tgccctccct
     3481 gtggatgcca cctctgatga agtcaggaag aacctgatgg acatgttcag ggacaggcag
     3541 gccttctctg aacacacctg gaagatgctc ctgtctgtgt gcagatcctg ggctgcctgg
     3601 tgcaagctga actgataccc agctttcttg tacaaagttg gcattataag aaagcattgc
     3661 ttatcaattt gttgcaacga acaggtcact atcagtcaaa ataaaatcat tatttgccat
     3721 ccagctgata tcccctatag tgagtcgtat tacatggtca tagctgtttc ctggcagctc
     3781 tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat atcatcatga
     3841 acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcaa
     3901 cgggaaacgt cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa
     3961 tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta tgggaagccc
     4021 gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat
     4081 gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt
     4141 atccgtactc ctgatgatgc atggttactc accactgcga tccccggaaa aacagcattc
     4201 caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc
     4261 ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt
     4321 cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat
     4381 gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa acttttgcca
     4441 ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac
     4501 gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag
     4561 gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt
     4621 tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc
     4681 gatgagtttt tctaatcaga attggttaat tggttgtaac actggcagag cattacgctg
     4741 acttgacggg acggcgcaag ctcatgacca aaatccctta acgtgagtta cgcgtcgttc
     4801 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     4861 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     4921 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     4981 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     5041 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     5101 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     5161 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     5221 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     5281 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     5341 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     5401 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     5461 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     5521 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     5581 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     5641 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     5701 agtgagcgca acgcaattaa tacgcgtacc gctagccagg aagagtttgt agaaacgcaa
     5761 aaaggccatc cgtcaggatg gccttctgct tagtttgatg cctggcagtt tatggcgggc
     5821 gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt caaatccgct cccggcggat
     5881 ttgtcctact caggagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt
     5941 ccgactgagc ctttcgtttt atttgatgcc tggcagttcc ctactctcgc gttaacgcta
     6001 gcatggatgt tttcccagtc acgacgttgt aaaacgacgg ccagtcttaa gctcgggccc
//
