LOCUS       sequence-335626-        4393 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  For light regulated release of transferrin receptor (TfR) from the 
            endoplasmic reticulum. Driven by CMV promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4393)
  AUTHORS   Bourke AM, Schwartz SL, Bowen AB, Kleinjan MS, Winborn CS, Kareemo 
            DJ, Gutnick A, Schwarz TL, Kennedy MJ
  TITLE     zapERtrap: A light-regulated ER release system reveals unexpected 
            neuronal trafficking pathways.
  JOURNAL   J Cell Biol. 2021 Sep 6;220(9). pii: 212461. doi: 
            10.1083/jcb.202103186. Epub 2021 Jul 9.
  PUBMED    34241635
REFERENCE   2  (bases 1 to 4393)
  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"; journalName: "J Cell 
            Biol."; date: "2021-09-6"; volume: "220(9). pii: 212461. doi"; 
            pages: " 10.1083/jcb.202103186. Epub 2021 Jul 9"
COMMENT     Sequence Label: TfR-mCh-DHFR
FEATURES             Location/Qualifiers
     source          1..4393
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        33..84
                     /label=CMVd2 promoter
                     /note="truncated human cytomegalovirus (CMV) immediate 
                     early promoter that yields moderately reduced expression 
                     relative to the full-length promoter"
     primer_bind     35..55
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     81..105
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        315..333
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     315..332
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     346..365
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             complement(1362..1373)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     regulatory      2718..2727
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2724..3428
                     /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
                     EGRHSTGGMDELY"
     primer_bind     complement(2869..2887)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3137..3156
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             3456..3932
                     /codon_start=1
                     /gene="folA"
                     /product="E. coli dihydrofolate reductase"
                     /label=ecDHFR
                     /translation="ISLIAALAVDRVIGMENAMPWNLPADLAWFKRNTLNKPVIMGRHT
                     WESIGRPLPGRKNIILSSQPGTDDRVTWVKSVDEAIAACGDVPEIMVIGGGRVYEQFLP
                     KAQKLYLTHIDAEVEGDTHFPDYEPDDWESVFSEFHDADAQNSHSYCFEILERR"
     regulatory      3970..3979
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3983..4117
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4020..4039)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4074..4093
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     enhancer        4186..4374
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4209..4229)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 agcatgcaaa tgtcgtacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
       61 tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcgcctg gagacgccat
      121 ccacgctgtt ttgacctcca tagaagacac cgggaccgat ccagcctccg cggccgggaa
      181 cggtgcattg gaacgcggat tccccgtgcc aagagtgacg taagtaccgc ctatagagtc
      241 tataggccca cccccttggc ttcgttagaa cgcggctaca attaatacat aaccttatgt
      301 atcatacaca tacgatttag gtgacactat agaataacat ccactttgcc tttctctcca
      361 caggtgtcca ctcccaggtc caactgcacc tcggttctat cgattgaatt ccccggggat
      421 ccaccatgga tcaagctaga tcagcattct ctaacttgtt tggtggagaa ccattgtcat
      481 atacccggtt cagcctggct cggcaagtag atggcgataa cagtcatgtg gagatgaaac
      541 ttgctgtaga tgaagaagaa aatgctgaca ataacacaaa ggccaatgtc acaaaaccaa
      601 aaaggtgtag tggaagtatc tgctatggga ctattgctgt gatcgtcttt ttcttgattg
      661 gatttatgat tggctacttg ggctattgta aaggggtaga accaaaaact gagtgtgaga
      721 gactggcagg aaccgagtct ccagtgaggg aggagccagg agaggacttc cctgcagcac
      781 gtcgcttata ttgggatgac ctgaagagaa agttgtcgga gaaactggac agcacagact
      841 tcaccggcac catcaagctg ctgaatgaaa attcatatgt ccctcgtgag gctggatctc
      901 aaaaagatga aaatcttgcg ttgtatgttg aaaatcaatt tcgtgaattt aaactcagca
      961 aagtctggcg tgatcaacat tttgttaaga ttcaggtcaa agacagcgct caaaactcgg
     1021 tgatcatagt tgataagaac ggtagacttg tttacctggt ggagaatcct gggggttatg
     1081 tggcgtatag taaggctgca acagttactg gtaaactggt ccatgctaat tttggtacta
     1141 aaaaagattt tgaggattta tacactcctg tgaatggatc tatagtgatt gtcagagcag
     1201 ggaaaatcac ctttgcagaa aaggttgcaa atgctgaaag cttaaatgca attggtgtgt
     1261 tgatatacat ggaccagact aaatttccca ttgttaacgc agaactttca ttctttggac
     1321 atgctcatct ggggacaggt gacccttaca cacctggatt cccttccttc aatcacactc
     1381 agtttccacc atctcggtca tcaggattgc ctaatatacc tgtccagaca atctccagag
     1441 ctgctgcaga aaagctgttt gggaatatgg aaggagactg tccctctgac tggaaaacag
     1501 actctacatg taggatggta acctcagaaa gcaagaatgt gaagctcact gtgagcaatg
     1561 tgctgaaaga gataaaaatt cttaacatct ttggagttat taaaggcttt gtagaaccag
     1621 atcactatgt tgtagttggg gcccagagag atgcatgggg ccctggagct gcaaaatccg
     1681 gtgtaggcac agctctccta ttgaaacttg cccagatgtt ctcagatatg gtcttaaaag
     1741 atgggtttca gcccagcaga agcattatct ttgccagttg gagtgctgga gactttggat
     1801 cggttggtgc cactgaatgg ctagagggat acctttcgtc cctgcattta aaggctttca
     1861 cttatattaa tctggataaa gcggttcttg gtaccagcaa cttcaaggtt tctgccagcc
     1921 cactgttgta tacgcttatt gagaaaacaa tgcaaaatgt gaagcatccg gttactgggc
     1981 aatttctata tcaggacagc aactgggcca gcaaagttga gaaactcact ttagacaatg
     2041 ctgctttccc tttccttgca tattctggaa tcccagcagt ttctttctgt ttttgcgagg
     2101 acacagatta tccttatttg ggtaccacca tggacaccta taaggaactg attgagagga
     2161 ttcctgagtt gaacaaagtg gcacgagcag ctgcagaggt cgctggtcag ttcgtgatta
     2221 aactaaccca tgatgttgaa ttgaacctgg actatgagag gtacaacagc caactgcttt
     2281 catttgtgag ggatctgaac caatacagag cagacataaa ggaaatgggc ctgagtttac
     2341 agtggctgta ttctgctcgt ggagacttct tccgtgctac ttccagacta acaacagatt
     2401 tcgggaatgc tgagaaaaca gacagatttg tcatgaagaa actcaatgat cgtgtcatga
     2461 gagtggagta tcacttcctc tctccctacg tatctccaaa agagtctcct ttccgacatg
     2521 tcttctgggg ctccggctct cacacgctgc cagctttact ggagaacttg aaactgcgta
     2581 aacaaaataa cggtgctttt aatgaaacgc tgttcagaaa ccagttggct ctagctactt
     2641 ggactattca gggagctgca aatgccctct ctggtgacgt ttgggacatt gacaatgagt
     2701 ttggggatcc accggtcgcc accatggtga gcaagggcga ggaggataac atggccatca
     2761 tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg
     2821 agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg
     2881 tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg
     2941 gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc
     3001 ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga
     3061 cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca
     3121 acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg
     3181 agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga
     3241 aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc
     3301 agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact
     3361 acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg
     3421 agctgtacag tgctggtagt gctggtagtg ctggtatcag tctgattgcg gcgttagcgg
     3481 tagatcgcgt tatcggcatg gaaaacgcca tgccgtggaa cctgcctgcc gatctcgcct
     3541 ggtttaaacg caacacctta aataaacccg tgattatggg ccgccatacc tgggaatcaa
     3601 tcggtcgtcc gttgccagga cgcaaaaata ttatcctcag cagtcaaccg ggtacggacg
     3661 atcgcgtaac gtgggtgaag tcggtggatg aagccatcgc ggcgtgtggt gacgtaccag
     3721 aaatcatggt gattggcggc ggtcgcgttt atgaacagtt cttgccaaaa gcgcaaaaac
     3781 tgtatctgac gcatatcgac gcagaagtgg aaggcgacac ccatttcccg gattacgagc
     3841 cggatgactg ggaatcggta ttcagcgaat tccacgatgc tgatgcgcag aactctcaca
     3901 gctattgctt tgagattctg gagcggcggt aaagcggccg cgactctaga gtcgacctgc
     3961 agaagcttgg ccgccatggc ccaacttgtt tattgcagct tataatggtt acaaataaag
     4021 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     4081 gtccaaactc atcaatgtat cttatcatgt ctggatcggg aattaattcg gcgcagcacc
     4141 atggcctgaa ataacctctg aaagaggaac ttggttaggt accttctgag gcggaaagaa
     4201 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcagcaga
     4261 agtatgcaaa gcatgcatct catttagtca gcaaccagtg tggaaagtcc ccaggctccc
     4321 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaccatag tcccgcccct
     4381 aactccgccc atc
//
