LOCUS       pTKI-Tyr_GFP-SUM        6852 bp    DNA     circular SYN 18-SEP-2023
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6852)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 1, 2025 from SnapGene 8.0.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6852
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          162..187
                     /mol_type="other DNA"
                     /note="color: #ffffff"
                     /organism="synthetic DNA construct"
     primer_bind     complement(135..183)
                     /label=pTet-sfGFP_gibR
                     /note="color: black; sequence: 
                     AGTCCGGAACGTCGTACGGGTACATATGGATACCGCCTGAGTCAGTACT"
     CDS             159..188
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="color: #339966"
                     /translation="MYPYDVPDYA"
     primer_bind     165..212
                     /label=pTet-sfGFP_gibF
                     /note="color: black; sequence: 
                     CCGTACGACGTTCCGGACTACGCCCGTAAAGGCGAAGAGCTGTTCACT"
     CDS             189..899
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=sfGFP
                     /note="color: #05fd14"
                     /translation="RKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDGT
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
                     FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(874..917)
                     /label=sfGFP-SUMO_gibR
                     /note="color: black; sequence: 
                     cgagtcCATCAATTGGCCTTTGTACAGTTCATCCATGCCATGCG"
     primer_bind     891..929
                     /label=sfGFP-SUMO_gibF
                     /note="color: black; sequence: 
                     CTGTACAAAGGCCAATTGATGgactcggaggtcaaccag"
     CDS             909..1199
                     /codon_start=1
                     /locus_tag="SUMO_myc_opt"
                     /label=SUMO
                     /label=SUMO_myc_opt
                     /note="color: #ff9900"
                     /translation="MDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPLR
                     RLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAYREQIGG"
     primer_bind     complement(1183..1227)
                     /label=SUMO-Tyr_mCherry_gibR
                     /note="color: black; sequence: 
                     TgGCTGTGCCAGATTGGACGCGCAGGCTgccaccgatctgctcgc"
     CDS             1202..1228
                     /codon_start=1
                     /locus_tag="Nend tag"
                     /label=Nend tag
                     /note="color: #c0c0c0"
                     /translation="LRVQSGTA"
     primer_bind     1208..1260
                     /label=SUMO-Nend_mCherry_gibF
                     /note="color: black; sequence: 
                     CGTCCAATCTGGCACAGCcAGCCTGCAGGtctcgaagggggaagaggacaaca"
     CDS             1237..1940
                     /codon_start=3
                     /label=mCherry
                     /note="color: #ff0000"
                     /translation="SKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGTQ
                     TAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFED
                     GGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKGE
                     IKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAEG
                     RHSTGGMDELYK"
     primer_bind     complement(1936..1988)
                     /label=mCherry-pTet_gibR
                     /note="color: black; sequence: 
                     CCTTAAGGCGGCCGCTTACAGATCCTCTTCTGAGATGAGTTTTTGTTCtttgt"
     CDS             1941..1970
                     /codon_start=1
                     /locus_tag="Myc tag"
                     /label=Myc tag
                     /note="color: #0080ff"
                     /translation="EQKLISEEDL"
     primer_bind     1969..2010
                     /label=mCherry-pTet_gibF
                     /note="color: black; sequence: 
                     TGTAAGCGGCCGCCTTAAGGCGGCCGCCTTAAAAAAAAAAGC"
     primer_bind     1981..1995
                     /label=mCherry-pTet_gibR
                     /note="color: black; sequence: 
                     CCTTAAGGCGGCCGCTTACAGATCCTCTTCTGAGATGAGTTTTTGTTCtttgt"
     rep_origin      complement(2140..2728)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     primer_bind     complement(2280..2299)
                     /label=pLJR962_seqR
                     /note="color: black; sequence: GGACAGGTATCCGGTAAGCG"
     terminator      2847..2874
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
                     /note="color: #ffffff"
     CDS             3264..3887
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /note="color: #993366"
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     CDS             4603..5601
                     /codon_start=1
                     /label=mycobacterial integrase
                     /note="color: #33cccc"
                     /translation="MDAEAWLAGEKRLIEMETWTPPQDRAKKAAASAITLEEYTRKWLV
                     ERDLADGTRDLYSGHAERRIYPVLGEVAVTEMTPALVRAWWAGMGRKHPTARRHAYNVL
                     RAVMNTAVEDKLIAENPCRIEQKAADERDVEALTPEELDIVAAEIFVHYRIAAYILAWT
                     SLRFGELIELRRKDIVDDGMTMKLRVRRGASRVGNKIVVGNAKTVRSKRPVTVPPHVAE
                     MIRAHMKDRTKMNKGPEAFLVTTTQGNRLSKSAFTKSLKRGYAKIGRPELRIHDLRAVG
                     ATFAAQAGATTKELMARLGHTTPRMAMKYQMASEARDEAIAEAMSKLAKTS"
     CDS             5901..6716
                     /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="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPHAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHNLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     6754..6773
                     /label=pLJR962_seqF
                     /note="color: black; sequence: TTACGCTGACTTGACGGGAC"
ORIGIN
        1 cgtggcaaag caaaagttca aaatcaccaa ctggtccacc tacaacaaag ctctcaccaa
       61 ccgtggtctg accagggaaa atagccctct gacctgggat ttgcgatccc tatcagtgat
      121 agatataatc tgggagtact gactcaggcg gtatccatat gtacccgtac gacgttccgg
      181 actacgcccg taaaggcgaa gagctgttca ctggtgtcgt ccctattctg gtggaactgg
      241 atggtgatgt caacggtcat aagttttccg tgcgtggcga gggtgaaggt gacgcaacta
      301 atggtaaact gacgctgaag ttcatctgta ctactggtaa actgccggta ccttggccga
      361 ctctggtaac gacgctgact tatggtgttc agtgctttgc tcgttatccg gaccatatga
      421 agcagcatga cttcttcaag tccgccatgc cggaaggcta tgtgcaggaa cgcacgattt
      481 cctttaagga tgacggcacg tacaaaacgc gtgcggaagt gaaatttgaa ggcgataccc
      541 tggtaaaccg cattgagctg aaaggcattg actttaaaga agacggcaat atcctgggcc
      601 ataagctgga atacaatttt aacagccaca atgtttacat caccgccgat aaacaaaaaa
      661 atggcattaa agcgaatttt aaaattcgcc acaacgtgga ggatggcagc gtgcagctgg
      721 ctgatcacta ccagcaaaac actccaatcg gtgatggtcc tgttctgctg ccagacaatc
      781 actatctgag cacgcaaagc gttctgtcta aagatccgaa cgagaaacgc gatcatatgg
      841 ttctgctgga gttcgtaacc gcagcgggca tcacgcatgg catggatgaa ctgtacaaag
      901 gccaattgat ggactcggag gtcaaccagg aggccaagcc cgaggtcaaa ccggaagtga
      961 agccggagac ccacatcaac ctgaaagtga gcgacggtag ctcggagatc ttcttcaaga
     1021 tcaagaagac caccccgctg cgccggctga tggaagcctt cgcgaagcgc caaggcaaag
     1081 agatggactc gctccgcttc ctctacgacg gtatccgcat ccaggccgac caaacccccg
     1141 aggatctgga catggaggac aatgacatca tcgaagctta ccgcgagcag atcggtggct
     1201 acctgcgcgt ccaatctggc acagccagcc tgcaggtctc gaagggggaa gaggacaaca
     1261 tggccatcat caaggaattc atgcggttca aggtgcacat ggagggcagc gtcaatgggc
     1321 atgagttcga gatcgagggg gaaggtgagg ggcggccgta cgaaggcacc cagacggcca
     1381 agctcaaggt cacgaagggc ggcccgctcc cgttcgcctg ggacatcctg agcccgcagt
     1441 tcatgtacgg ctcgaaggcc tacgtgaagc atccggccga catcccggac tacctcaagc
     1501 tctcgttccc cgaaggcttc aagtgggagc gtgtcatgaa cttcgaggac ggcggtgtgg
     1561 tcacggtcac ccaggactcc tcgctgcaag atggggaatt catctacaag gtcaagctcc
     1621 gggggaccaa cttcccctcg gacggtccgg tgatgcagaa gaagaccatg ggttgggaag
     1681 ccagctccga gcgcatgtac ccggaggacg gtgccctgaa gggggaaatc aagcagcggc
     1741 tcaagctgaa ggacggtggt cattacgacg ccgaggtgaa gaccacctac aaggccaaaa
     1801 agccggtcca actgcccggc gcctacaacg tgaacatcaa actggatatc acctcgcaca
     1861 acgaggacta caccatcgtc gagcagtacg agcgcgccga gggtcgccac agcaccggtg
     1921 gcatggatga actgtacaaa gaacaaaaac tcatctcaga agaggatctg taagcggccg
     1981 ccttaaggcg gccgccttaa aaaaaaaagc cccgcgattg cggggctttt tttttttaat
     2041 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     2101 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     2161 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     2221 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     2281 gcttaccgga tacctgtccg cctttctccc ctcgggaagc gtggcgcttt ctcatagctc
     2341 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     2401 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     2461 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     2521 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     2581 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     2641 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     2701 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     2761 cgctcatgga acaataaacg aaaggctcag tcgaaagact gggcctttcg tttatctgtt
     2821 gtttgaaagc cgctgattag ctaagcagaa ggccatcctg acggatggcc tttttctaga
     2881 aaaaaaaaaa gcgccgcaac tgcggcgctt tttttttttt aattaaagaa atattggatc
     2941 gtcggcaccg tcacggccgt gggaggcggc acgatccgcg acgtgatgat cggccgcatc
     3001 cccacggtgc tgcgcagtga gctctacgcc atcccggcgt tgatctgtgc gttcgcacgc
     3061 acaggcccgg tgtgagaagg gtctctgacg agcgggagaa cccacccggg gtgggcgagt
     3121 ttgtcctgcg tgtgctcggt cgagtaggct ctgggagtac ccgtgtgtac gaccagcacg
     3181 gcatacatca tttcgacgcc gagagattcg ccgcccgaaa tgagcacgat ccgcatggta
     3241 ccatcaggag gaatcacttc gcaatgtcta gattagataa aagtaaagtg attaacagcg
     3301 cattagagct gcttaatgag gtcggaatcg aaggtttaac aacccgtaaa ctcgcccaga
     3361 agcttggtgt agagcagcct acattgtatt ggcatgtaaa aaataagcgg gccctgctcg
     3421 acgccttagc cattgagatg ttagataggc accatactca cttttgccct ttagaagggg
     3481 aaagctggca agatttttta cgtaataacg ctaaaagttt tagatgtgct ttactaagtc
     3541 atcgcgatgg agcaaaagta catttaggta cacggcctac agaaaaacag tatgaaactc
     3601 tcgaaaatca attagccttt ttatgccaac aaggtttttc actagagaac gcgttatatg
     3661 cactcagcgc tgtggggcat tttaccttag gttgcgtatt ggaagatcaa gagcatcaag
     3721 tcgctaaaga agaaagggaa acacctacta ctgatagtat gccgccatta ttacgacaag
     3781 ctatcgaatt atttgatcac caaggtgcag agccagcctt cttattcggc cttgaattga
     3841 tcatctgcgg attagaaaaa caacttaaat gtgaaagtgg gtcttaatga ccatgggcta
     3901 gcgctttagc taattaattt aaaataaaaa aggggacctc tagggtcccc aattaaatag
     3961 ttaacactag tattaaaaaa aaaagccccg cgattgcggg gctttttttt ttcgacggcg
     4021 agggttccga ccgctgcaac tcccggtgca accttgtccc ggtctattct cttcactgca
     4081 ccagctccaa tctggtgtga atgcccctcg tctgttcgcg caggcggggg gctctattcg
     4141 tttgtcagca tcgaaagtag ccagatcagg gatgcgttgc aaccgcgtat gcccaggtca
     4201 gaagagtcgc acaagagttg cagacccctg gaaagaaaaa tggccagagg gcgaaaacac
     4261 cctctgacca gcggagcggg cgacgggaat cgaacccgcg tagctagttt ggaagaatgg
     4321 gtgtctgccg accacatata tgggccggtc aagataggtt tttaccccct ctcggctgca
     4381 tcctctaagt ggaaagaaat tgcaggtcgt agaagcgcgt tgaagcctga gagttgcaca
     4441 ggagttgcaa cccggtagcc ttgttcacga cgagaggaga cctagttggc acgtcgcgga
     4501 tggggatcgc tgaagactca gcgcagcggg aggatccaag cctcatacgt caacccgcag
     4561 gacggtgtga ggtactacgc gctgcagacc tacgacaaca agatggacgc cgaagcctgg
     4621 ctcgcgggcg agaagcggct catcgagatg gagacctgga cccctccaca ggaccgggcg
     4681 aagaaggcag ccgccagcgc catcacgctg gaggagtaca cccggaagtg gctcgtggag
     4741 cgcgacctcg cagacggcac cagggatctg tacagcgggc acgcggagcg ccgcatctac
     4801 ccggtgctag gtgaagtggc ggtcacagag atgacgccag ctctggtgcg tgcgtggtgg
     4861 gccgggatgg gtaggaagca cccgactgcc cgccggcatg cctacaacgt cctccgggcg
     4921 gtgatgaaca cagcggtcga ggacaagctg atcgcagaga acccgtgccg gatcgagcag
     4981 aaggcagccg atgagcgcga cgtagaggcg ctgacgcctg aggagctgga catcgtcgcc
     5041 gctgagatct tcgtgcacta ccggatcgcg gcatacatcc tggcgtggac gagcctccgg
     5101 ttcggagagc tgatcgagct tcgccgcaag gacatcgtgg acgacggcat gacgatgaag
     5161 ctccgggtgc gccgtggcgc ttcccgcgtg gggaacaaga tcgtcgttgg caacgccaag
     5221 accgtccggt cgaagcgtcc tgtgacggtt ccgcctcacg tcgcggagat gatccgagcg
     5281 cacatgaagg accgtacgaa gatgaacaag ggccccgagg cattcctggt gaccacgacg
     5341 cagggcaacc ggctgtcgaa gtccgcgttc accaagtcgc tgaagcgtgg ctacgccaag
     5401 atcggtcggc cggaactccg catccacgac ctccgcgctg tcggcgctac gttcgccgct
     5461 caggcaggtg cgacgaccaa ggagctgatg gcccgcctcg gtcacacgac tcctaggatg
     5521 gcgatgaagt accagatggc gtctgaggcc cgcgacgagg ctatcgctga ggcgatgtcc
     5581 aagctggcca agacctcctg aaacgcaaaa agcccccctc ccaaggacac tgagtcctaa
     5641 agaggggggt ttcttgtcag tacgcgaaga accacgcctg gccgcgagcg ccagcaccgc
     5701 cgctctgtgc ggagacctgg gcaccagccc cgccgccgcc aggagcattg ccgttcccgc
     5761 cagctagcca cctgacgtcg gggggggggg aaagccacgt tgtgtctcaa aatctctgat
     5821 gttacattgc acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa
     5881 acagtaatac aaggggtgtt atgagccata ttcaacggga aacgtcttgc tcgaggccgc
     5941 gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg
     6001 ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc ccatgcgcca gagttgtttc
     6061 tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact
     6121 ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg
     6181 catggttact caccactgcg atccccggga aaacagcatt ccaggtatta gaagaatatc
     6241 ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga
     6301 ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgcctcgct caggcgcaat
     6361 cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc
     6421 ctgttgaaca agtctggaaa gaaatgcata atcttttgcc attctcaccg gattcagtcg
     6481 tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt
     6541 gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga
     6601 actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg
     6661 ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag
     6721 aattggttaa ttggttgtaa cactggcaga gcattacgct gacttgacgg gacggcggct
     6781 ttgttgaata aatcgaactt ttgctgagtt gaaggatcag atcacgcatc ttcccgacaa
     6841 cgcagaccgt tc
//
