LOCUS       sequence-134730-        7747 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses KLC1-mCherry-eDHFR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7747)
  AUTHORS   Ballister ER, Ayloo S, Chenoweth DM, Lampson MA, Holzbaur EL
  TITLE     Optogenetic control of organelle transport using a photocaged 
            chemical inducer of dimerization.
  JOURNAL   Curr Biol. 2015 May 18;25(10):R407-8. doi: 
            10.1016/j.cub.2015.03.056.
  PUBMED    25989077
REFERENCE   2  (bases 1 to 7747)
  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: 
            "10.1016/j.cub.2015.03"; journalName: "Curr Biol"; date: 
            "2015-05-18- 18"; volume: "25"; issue: "10"; pages: "R407-8"
COMMENT     Sequence Label: pERB257
FEATURES             Location/Qualifiers
     source          1..7747
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    52..100
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     protein_bind    312..345
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     CDS             350..949
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(350..369)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     846..866
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(1085..1102)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1091..1202
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        1347..1650
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1652..1929
                     /label=chicken beta-actin promoter
     intron          1930..2946
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2870..2892
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2954..2973
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             3556..4263
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3701..3719)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3969..3988
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             4282..4755
                     /codon_start=1
                     /gene="folA"
                     /product="E. coli dihydrofolate reductase"
                     /label=ecDHFR
                     /translation="ISLIAALAVDRVIGMENAMPWNLPADLAWFKRNTLNKPVIMGRHT
                     WESIGRPLPGRKNIILSSQPGTDDRVTWVKSVDEAIAACGDVPEIMVIGGGRVYEQFLP
                     KAQKLYLTHIDAEVEGDTHFPDYEPDDWESVFSEFHDADAQNSHSYCFEILERR"
     CDS             4756..4785
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(4892..4911)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4957..5012
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5011..5030)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     protein_bind    5375..5408
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    5421..5542
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5458..5477)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5512..5531
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5561..5581)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5561..5579)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5600..5616)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5600..5616)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5613..5635)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5624..5640
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5648..5654,5655..5672,5673..5678))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5693..5714
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5831..5848)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6002..6590)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6082..6101)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6761..7552,7553..7621))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7384..7403
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7622..7726)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aagcttaata aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
       61 aaagacagaa taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
      121 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
      181 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
      241 gtcggggcgg caggccctgc catagccact ggccccgtgg gttagggacg gggtccccca
      301 ttcgagaata aataacttcg tataaagtat cctatacgaa gttatcacca tgaccgagta
      361 caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc cgggccgtac gcaccctcgc
      421 cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gacccggacc gccacatcga
      481 gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt
      541 gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc
      601 gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc
      661 cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt
      721 cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt
      781 gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc
      841 gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt
      901 gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgac gcccgcccca
      961 cgacccgcag cgcccgaccg aaaggagcgc acgaccccat ggctccgacc gaagccgacc
     1021 cgggcggccc cgccgacccc gcacccgccc ccgaggccca ccgactctag agctcgctga
     1081 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     1141 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     1201 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     1261 ggggaggatt gggaagacaa tagcactagt tattaatagt aatcaattac ggggtcatta
     1321 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
     1381 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
     1441 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
     1501 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
     1561 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     1621 atctacgtat tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact
     1681 ctccccatct cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt
     1741 tgtgcagcga tgggggcggg gggggggggg gggcgcgcgc caggcggggc ggggcggggc
     1801 gaggggcggg gcggggcgag gcggagaggt gcggcggcag ccaatcagag cggcgcgctc
     1861 cgaaagtttc cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg
     1921 cggcgggcgg gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg
     1981 cgccgcccgc cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc
     2041 ccttctcctc cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg
     2101 ctgcgtgaaa gccttgaggg gctccgggag ggccctttgt gcggggggag cggctcgggg
     2161 ggtgcgtgcg tgtgtgtgtg cgtggggagc gccgcgtgcg gctccgcgct gcccggcggc
     2221 tgtgagcgct gcgggcgcgg cgcggggctt tgtgcgctcc gcagtgtgcg cgaggggagc
     2281 gcggccgggg gcggtgcccc gcggtgcggg gggggctgcg aggggaacaa aggctgcgtg
     2341 cggggtgtgt gcgtgggggg gtgagcaggg ggtgtgggcg cgtcggtcgg gctgcaaccc
     2401 cccctgcacc cccctccccg agttgctgag cacggcccgg cttcgggtgc ggggctccgt
     2461 acggggcgtg gcgcggggct cgccgtgccg ggcggggggt ggcggcaggt gggggtgccg
     2521 ggcggggcgg ggccgcctcg ggccggggag ggctcggggg aggggcgcgg cggcccccgg
     2581 agcgccggcg gctgtcgagg cgcggcgagc cgcagccatt gccttttatg gtaatcgtgc
     2641 gagagggcgc agggacttcc tttgtcccaa atctgtgcgg agccgaaatc tgggaggcgc
     2701 cgccgcaccc cctctagcgg gcgcggggcg aagcggtgcg gcgccggcag gaaggaaatg
     2761 ggcggggagg gccttcgtgc gtcgccgcgc cgccgtcccc ttctccctct ccagcctcgg
     2821 ggctgtccgc ggggggacgg ctgccttcgg gggggacggg gcagggcggg gttcggcttc
     2881 tggcgtgtga ccggcggctc tagagcctct gctaaccatg ttcatgcctt cttctttttc
     2941 ctacagctcc tgggcaacgt gctggttatt gtgctgtctc atcattttgg caaagaattg
     3001 ctcgaccacc ggcgcgccat gcatgacaac atgtccacaa tggtgtacat gaaggaagag
     3061 aagctggaga agcttacgca ggatgaaatc atttctaaga ccaagcaggt gatccagggg
     3121 ctggaggccc tgaagaatga gcacaactcc atcctgcaga gtctgctgga gacgctgaag
     3181 tgcttgaaga aggacgatga gagcaacctg gtggaggaga agtccagcat gatccgcaag
     3241 tccttggaga tgctggagct cggcctgagc gaggcgcagg tgatgatggc cctgtctaat
     3301 cacctgaatg ccgtggagtc cgagaagcag aagctccgag ctcaggttcg acgtctgtgc
     3361 caggagaacc agtggctgcg ggatgaactg gccaacacac agcagaagtt acagaagagc
     3421 gagcagtccg tggctcagct ggaggaggag aagaagcacc tggagttcat gaaccagctg
     3481 aagaaatacg acgacgacat ctctccctcg gaagacaaag actctgattc ttccaaagag
     3541 ccgggaccgg tcaccatggt gagcaagggc gaggaggata acatggccat catcaaggag
     3601 ttcatgcgct tcaaggtgca catggagggc tccgtgaacg gccacgagtt cgagatcgag
     3661 ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag
     3721 ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc agttcatgta cggctccaag
     3781 gcctacgtga agcaccccgc cgacatcccc gactacttga agctgtcctt ccccgagggc
     3841 ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt gacccaggac
     3901 tcctccctgc aggacggcga gttcatctac aaggtgaagc tgcgcggcac caacttcccc
     3961 tccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctcctc cgagcggatg
     4021 taccccgagg acggcgccct gaagggcgag atcaagcaga ggctgaagct gaaggacggc
     4081 ggccactacg acgctgaggt caagaccacc tacaaggcca agaagcccgt gcagctgccc
     4141 ggcgcctaca acgtcaacat caagttggac atcacctccc acaacgagga ctacaccatc
     4201 gtggaacagt acgaacgcgc cgagggccgc cactccaccg gcggcatgga cgagctgtat
     4261 aaggccggcc tggagtctag aatcagtctg attgcggcgt tagcggtaga tcgcgttatc
     4321 ggcatggaaa acgccatgcc gtggaacctg cctgccgatc tcgcctggtt taaacgcaac
     4381 accttaaata aacccgtgat tatgggccgc catacctggg aatcaatcgg tcgtccgttg
     4441 ccaggacgca aaaatattat cctcagcagt caaccgggta cggacgatcg cgtaacgtgg
     4501 gtgaagtcgg tggatgaagc catcgcggcg tgtggtgacg taccagaaat catggtgatt
     4561 ggcggcggtc gcgtttatga acagttcttg ccaaaagcgc aaaaactgta tctgacgcat
     4621 atcgacgcag aagtggaagg cgacacccat ttcccggatt acgagccgga tgactgggaa
     4681 tcggtattca gcgaattcca cgatgctgat gcgcagaact ctcacagcta ttgctttgag
     4741 attctggagc ggcgggaaca aaaactcatc tcagaagagg atctttaatt aagcgctcga
     4801 ggcggccgca attcactcct caggtgcagg ctgcctatca gaaggtggtg gctggtgtgg
     4861 ccaatgccct ggctcacaaa taccactgag atctttttcc ctctgccaaa aattatgggg
     4921 acatcatgaa gccccttgag catctgactt ctggctaata aaggaaattt attttcattg
     4981 caatagtgtg ttggaatttt ttgtgtctct cactcggaag gacatatggg agggcaaatc
     5041 atttaaaaca tcagaatgag tatttggttt agagtttggc aacatatgcc catatgctgg
     5101 ctgccatgaa caaaggttgg ctataaagag gtcatcagta tatgaaacag ccccctgctg
     5161 tccattcctt attccataga aaagccttga cttgaggtta gatttttttt atattttgtt
     5221 ttgtgttatt tttttcttta acatccctaa aattttcctt acatgtttta ctagccagat
     5281 ttttcctcct ctcctgacta ctcccagtca tagctgtccc tcttctctta tgaagatccc
     5341 tcgacctgag cgtcgagcta gcatatggat ccatataact tcgtataatg tatgctatac
     5401 gaagttatta cgataccgtc aacttgttta ttgcagctta taatggttac aaataaagca
     5461 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     5521 ccaaactcat caatgtatct taggtaccca gcttttgttc cctttagtga gggttaattg
     5581 cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     5641 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     5701 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     5761 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     5821 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     5881 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     5941 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     6001 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     6061 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     6121 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     6181 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     6241 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     6301 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     6361 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     6421 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     6481 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     6541 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     6601 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     6661 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     6721 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     6781 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     6841 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     6901 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     6961 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     7021 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     7081 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     7141 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     7201 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     7261 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     7321 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     7381 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     7441 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     7501 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     7561 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     7621 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     7681 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     7741 aagtgcc
//
