LOCUS       pENTR221-Illusia        4845 bp DNA     circular SYN 11-JUN-2024
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 4845)
  AUTHORS   Ivaska laboratory
  TITLE     Direct Submission
  JOURNAL   Exported Jun 11, 2024 from SnapGene Viewer 7.2.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4845
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     terminator      225..252
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
                     /note="color: #ffffff"
     terminator      344..430
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
                     /note="color: #ffffff"
     primer_bind     494..510
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: RIGHT"
     protein_bind    526..625
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
                     /note="color: #31849b; direction: RIGHT"
     CDS             641..694
                     /codon_start=1
                     /label=LYN DOMAIN
                     /note="color: #993366"
                     /translation="MGCIKSKRKDNLNDDGVD"
     CDS             701..1414
                     /codon_start=1
                     /label=YPet
                     /note="color: #ffff00"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLLCTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALFKDPNEKRDHMVLL
                     EFLTAAGITEGMNELY"
     CDS             1421..1747
                     /codon_start=1
                     /product="EGSQFWVTVQRTEAAERCGLHGSYVLRVEAERLTLLTVGAQSQILEPLL
                     SWPYTLLRRYG
                     RDKVMFSFEAGRRCPSGPGTFTFQTAQGNDIFQAVETAIHRQKAQGKAG
                     
                     http://www.uniprot.org/uniprot/Q99704
                     "
                     /label=Dok1 PTB
                     /note="color: #339966"
                     /translation="EGSQFWVTVQRTEAAERCGLHGSYVLRVEAERLTLLTVGAQSQIL
                     EPLLSWPYTLLRRYGRDKVMFSFEAGRRCPSGPGTFTFQTAQGNDIFQAVETAIHRQKA
                     QGKAG"
     CDS             1754..2101
                     /codon_start=1
                     /label=Eevee Linker
                     /note="color: #a6acb3"
                     /translation="SAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGS
                     AGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGG
                     SAGGSAGGSAGG"
     CDS             2108..2188
                     /codon_start=1
                     /label=ITGB1 Cytoplasmic Domain (YYEE)
                     /note="color: #993366"
                     /translation="KGEEKPNVVTTVASKEIPNEGTDWKAN"
     CDS             join(2198..2200,2201..2203,2204..2914)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /note="This feature has 3 segments:
                        1: 2198 .. 2200 / #00ffff
                        2: 2201 .. 2203 / #00ffff / 1a
                        3: 2204 .. 2914 / #00ffff"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     protein_bind    complement(2927..3026)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
                     /note="color: #31849b; direction: LEFT"
     primer_bind     complement(3067..3083)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     CDS             3196..4005
                     /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"
                     /note="color: #ccffcc"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      4152..4740
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
ORIGIN
        1 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
       61 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
      121 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
      181 attaatacgc gtaccgctag ccaggaagag tttgtagaaa cgcaaaaagg ccatccgtca
      241 ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc
      301 ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg cggatttgtc ctactcagga
      361 gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtcttccgac tgagcctttc
      421 gttttatttg atgcctggca gttccctact ctcgcgttaa cgctagcatg gatgttttcc
      481 cagtcacgac gttgtaaaac gacggccagt cttaagctcg ggccccaaat aatgatttta
      541 ttttgactga tagtgacctg ttcgttgcaa caaattgatg agcaatgctt ttttataatg
      601 ccaactttgt acaaaaaagc aggctagcgt cgacgccacc atgggctgca tcaagagcaa
      661 gcgcaaggac aacctgaacg acgacggcgt ggactctaga atggtgagca agggcgagga
      721 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
      781 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagct
      841 tctatgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgggcta
      901 cggcctgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc
      961 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     1021 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     1081 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     1141 cagccacaac gtctatatca ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa
     1201 gatccgccac aacatcgagg acggcggcgt gcagctcgcc gaccactacc agcagaacac
     1261 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagct accagtccgc
     1321 cctgttcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcctgaccgc
     1381 cgccgggatc actgagggca tgaacgagct gtacctcgag gaaggttccc aattctgggt
     1441 aacggtgcag aggactgagg ccgccgagcg ctgtggcctg catggctcct acgtgctgag
     1501 ggtggaggct gaaaggctga ctctcctgac cgtgggggcc cagagtcaga tactggagcc
     1561 actcctgtcc tggccctaca ctctgttgcg tcgctatggc cgggacaagg tcatgttctc
     1621 tttcgaggcc ggccgccgct gcccctcagg ccctggaacc ttcaccttcc agacggcaca
     1681 gggaaatgac atcttccagg cagttgagac tgccatccac cggcagaagg cccagggaaa
     1741 ggccggaggt accagtgctg gtggtagtgc tggtggtagt gctggtggta gtgctggtgg
     1801 tagtgctggt ggttccggca gtgctggtgg tagtgctggt ggtagtacca gtgctggtgg
     1861 tagtgctggt ggtagtgctg gtggtagtgc tggtggtagt gctggtggtt ccggcagtgc
     1921 tggtggtagt gctggtggta gtaccagtgc tggtggtagt gctggtggta gtgctggtgg
     1981 tagtgctggt ggtagtgctg gtggttccgg cagtgctggt ggtagtgctg gtggtagtac
     2041 cagtgctggt ggtagtgctg gtggtagtgc tggtggtagt gctggtggta gtgctggtgg
     2101 ttccggaaaa ggagaggaaa agccgaatgt cgtgactaca gtagccagta aggaaattcc
     2161 taatgaaggt acggactgga aagccaatgg cggccgcatg gtgagcaagg gcgaggagct
     2221 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     2281 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
     2341 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgtcctgggg
     2401 cgtgcagtgc ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     2461 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     2521 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     2581 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actactttag
     2641 cgacaacgtc tatatcaccg ccgacaagca gaagaacggc atcaaggcca acttcaagat
     2701 ccgccacaac atcgaggacg gcggcgtgca gctcgccgac cactaccagc agaacacccc
     2761 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccaagct
     2821 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     2881 cgggatcact ctcggcatgg acgagctgta caagtaatga gaattcaccc agctttcttg
     2941 tacaaagttg gcattataag aaagcattgc ttatcaattt gttgcaacga acaggtcact
     3001 atcagtcaaa ataaaatcat tatttgccat ccagctgata tcccctatag tgagtcgtat
     3061 tacatggtca tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac
     3121 attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt
     3181 aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg attaaattcc
     3241 aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt
     3301 gcgacaatct atcgcttgta tgggaagccc gatgcgccag agttgtttct gaaacatggc
     3361 aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa
     3421 tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc
     3481 accactgcga tccccggaaa aacagcattc caggtattag aagaatatcc tgattcaggt
     3541 gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt
     3601 aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat
     3661 aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa
     3721 gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt cactcatggt
     3781 gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt
     3841 ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt
     3901 gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat
     3961 atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcaga attggttaat
     4021 tggttgtaac actggcagag cattacgctg acttgacggg acggcgcaag ctcatgacca
     4081 aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc
     4141 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     4201 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     4261 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     4321 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     4381 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     4441 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     4501 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     4561 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     4621 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     4681 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     4741 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     4801 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttacc
//
