LOCUS       sequence-339204-        8174 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For light regulated release of transferrin receptor (TfR) from the 
            endoplasmic reticulum. Driven by CMV promoter. Fused to oxidation 
            resistant version of GFP (moxGFP)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8174)
  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 8174)
  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-moxGFP-DHFR
FEATURES             Location/Qualifiers
     source          1..8174
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..22
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(37..42,43..60,61..67)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    75..91
                     /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)."
     primer_bind     80..102
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     99..115
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     99..115
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        144..523
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        524..727
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     678..698
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     724..748
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        958..976
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     958..975
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     989..1008
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             complement(2005..2016)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     regulatory      3361..3370
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3367..3369,3370..3372,3373..4080)
                     /codon_start=1
                     /product="monomeric GFP variant optimized for use in 
                     oxidizing environments such as the secretory pathway 
                     (Costantini et al., 2015)"
                     /label=moxGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFISTTGKLPVPWPTLVTTLTYGVQSFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELY"
     primer_bind     complement(3412..3433)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             4108..4584
                     /codon_start=1
                     /gene="folA"
                     /product="E. coli dihydrofolate reductase"
                     /label=ecDHFR
                     /translation="ISLIAALAVDRVIGMENAMPWNLPADLAWFKRNTLNKPVIMGRHT
                     WESIGRPLPGRKNIILSSQPGTDDRVTWVKSVDEAIAACGDVPEIMVIGGGRVYEQFLP
                     KAQKLYLTHIDAEVEGDTHFPDYEPDDWESVFSEFHDADAQNSHSYCFEILERR"
     regulatory      4604..4613
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    4617..4751
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4654..4673)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4708..4727
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        4820..5177
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4820..5011
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4843..4863)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5028..5163
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5197..5214)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5197..5213)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5206..5228)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5426..5881
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5513..5532)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5723..5744
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5898..5917)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6017..6039
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6077..6095)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6163..6267
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6268..6336,6337..7128)
                     /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     complement(6486..6505)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7299..7887
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7788..7807
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8041..8058
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
       61 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgacatgat
      121 tacgaattaa ttcgagctcg cccgacattg attattgact agttattaat agtaatcaat
      181 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
      241 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
      301 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
      361 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
      421 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
      481 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
      541 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
      601 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
      661 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
      721 cagagctcgt ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct
      781 ccatagaaga caccgggacc gatccagcct ccgcggccgg gaacggtgca ttggaacgcg
      841 gattccccgt gccaagagtg acgtaagtac cgcctataga gtctataggc ccaccccctt
      901 ggcttcgtta gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt
      961 taggtgacac tatagaataa catccacttt gcctttctct ccacaggtgt ccactcccag
     1021 gtccaactgc acctcggttc tatcgattga attccccggg gatccaccat ggatcaagct
     1081 agatcagcat tctctaactt gtttggtgga gaaccattgt catatacccg gttcagcctg
     1141 gctcggcaag tagatggcga taacagtcat gtggagatga aacttgctgt agatgaagaa
     1201 gaaaatgctg acaataacac aaaggccaat gtcacaaaac caaaaaggtg tagtggaagt
     1261 atctgctatg ggactattgc tgtgatcgtc tttttcttga ttggatttat gattggctac
     1321 ttgggctatt gtaaaggggt agaaccaaaa actgagtgtg agagactggc aggaaccgag
     1381 tctccagtga gggaggagcc aggagaggac ttccctgcag cacgtcgctt atattgggat
     1441 gacctgaaga gaaagttgtc ggagaaactg gacagcacag acttcaccag caccatcaag
     1501 ctgctgaatg aaaattcata tgtccctcgt gaggctggat ctcaaaaaga tgaaaatctt
     1561 gcgttgtatg ttgaaaatca atttcgtgaa tttaaactca gcaaagtctg gcgtgatcaa
     1621 cattttgtta agattcaggt caaagacagc gctcaaaact cggtgatcat agttgataag
     1681 aacggtagac ttgtttacct ggtggagaat cctgggggtt atgtggcgta tagtaaggct
     1741 gcaacagtta ctggtaaact ggtccatgct aattttggta ctaaaaaaga ttttgaggat
     1801 ttatacactc ctgtgaatgg atctatagtg attgtcagag cagggaaaat cacctttgca
     1861 gaaaaggttg caaatgctga aagcttaaat gcaattggtg tgttgatata catggaccag
     1921 actaaatttc ccattgttaa cgcagaactt tcattctttg gacatgctca tctggggaca
     1981 ggtgaccctt acacacctgg attcccttcc ttcaatcaca ctcagtttcc accatctcgg
     2041 tcatcaggat tgcctaatat acctgtccag acaatctcca gagctgctgc agaaaagctg
     2101 tttgggaata tggaaggaga ctgtccctct gactggaaaa cagactctac atgtaggatg
     2161 gtaacctcag aaagcaagaa tgtgaagctc actgtgagca atgtgctgaa agagataaaa
     2221 attcttaaca tctttggagt tattaaaggc tttgtagaac cagatcacta tgttgtagtt
     2281 ggggcccaga gagatgcatg gggccctgga gctgcaaaat ccggtgtagg cacagctctc
     2341 ctattgaaac ttgcccagat gttctcagat atggtcttaa aagatgggtt tcagcccagc
     2401 agaagcatta tctttgccag ttggagtgct ggagactttg gatcggttgg tgccactgaa
     2461 tggctagagg gatacctttc gtccctgcat ttaaaggctt tcacttatat taatctggat
     2521 aaagcggttc ttggtaccag caacttcaag gtttctgcca gcccactgtt gtatacgctt
     2581 attgagaaaa caatgcaaaa tgtgaagcat ccggttactg ggcaatttct atatcaggac
     2641 agcaactggg ccagcaaagt tgagaaactc actttagaca atgctgcttt ccctttcctt
     2701 gcatattctg gaatcccagc agtttctttc tgtttttgcg aggacacaga ttatccttat
     2761 ttgggtacca ccatggacac ctataaggaa ctgattgaga ggattcctga gttgaacaaa
     2821 gtggcacgag cagctgcaga ggtcgctggt cagttcgtga ttaaactaac ccatgatgtt
     2881 gaattgaacc tggactatga gaggtacaac agccaactgc tttcatttgt gagggatctg
     2941 aaccaataca gagcagacat aaaggaaatg ggcctgagtt tacagtggct gtattctgct
     3001 cgtggagact tcttccgtgc tacttccaga ctaacaacag atttcgggaa tgctgagaaa
     3061 acagacagat ttgtcatgaa gaaactcaat gatcgtgtca tgagagtgga gtatcacttc
     3121 ctctctccct acgtatctcc aaaagagtct cctttccgac atgtcttctg gggctccggc
     3181 tctcacacgc tgccagcttt actggagaac ttgaaactgc gtaaacaaaa taacggtgct
     3241 tttaatgaaa cgctgttcag aaaccagttg gctctagcta cttggactat tcagggagct
     3301 gcaaatgccc tctctggtga cgtttgggac attgacaatg agtttgggga tccaccggtc
     3361 gccaccatgg tgtccaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     3421 ctggacggcg acgtaaacgg ccacaagttc tccgtgcggg gcgagggcga gggcgatgcc
     3481 accaacggca agctgaccct gaagttcatc agcaccaccg gcaagctgcc cgtgccctgg
     3541 cccaccctcg tgaccaccct gacctacggc gtgcagagct tctcccgcta ccccgaccac
     3601 atgaagcgcc acgacttctt caagagcgcc atgcccgaag gctacgtcca ggagcgcacc
     3661 atctccttca aggacgacgg cacctacaag acccgcgccg aggtgaagtt cgagggcgac
     3721 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     3781 gggcacaagc tggagtacaa cttcaactcc cacaacgtct atatcaccgc cgacaagcag
     3841 aagaacggca tcaaggccaa cttcaagatc cgccacaacg tggaggacgg ctccgtgcag
     3901 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     3961 aaccactacc tgtccaccca gtccaagctg tccaaagacc ccaacgagaa gcgcgatcac
     4021 atggtccttc tggaattcgt gaccgccgcc gggatcactc acggcatgga cgagctgtac
     4081 agtgctggta gtgctggtag tgctggtatc agtctgattg cggcgttagc ggtagatcgc
     4141 gttatcggca tggaaaacgc catgccgtgg aacctgcctg ccgatctcgc ctggtttaaa
     4201 cgcaacacct taaataaacc cgtgattatg ggccgccata cctgggaatc aatcggtcgt
     4261 ccgttgccag gacgcaaaaa tattatcctc agcagtcaac cgggtacgga cgatcgcgta
     4321 acgtgggtga agtcggtgga tgaagccatc gcggcgtgtg gtgacgtacc agaaatcatg
     4381 gtgattggcg gcggtcgcgt ttatgaacag ttcttgccaa aagcgcaaaa actgtatctg
     4441 acgcatatcg acgcagaagt ggaaggcgac acccatttcc cggattacga gccggatgac
     4501 tgggaatcgg tattcagcga attccacgat gctgatgcgc agaactctca cagctattgc
     4561 tttgagattc tggagcggcg gtaagtcgac ctgcagaagc ttggccgcca tggcccaact
     4621 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     4681 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     4741 atgtctggat cgggaattaa ttcggcgcag caccatggcc tgaaataacc tctgaaagag
     4801 gaacttggtt aggtaccttc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
     4861 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4921 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4981 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     5041 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     5101 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     5161 gcctaggctt ttgcaaaaag ctgttaacag cttggcactg gccgtcgttt tacaacgtcg
     5221 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
     5281 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     5341 gaatggcgaa tggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     5401 ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt
     5461 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     5521 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     5581 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     5641 ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     5701 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     5761 atctcgggct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     5821 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt
     5881 ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     5941 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     6001 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     6061 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     6121 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     6181 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     6241 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     6301 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     6361 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     6421 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     6481 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     6541 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     6601 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     6661 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     6721 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     6781 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     6841 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     6901 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     6961 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     7021 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     7081 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     7141 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     7201 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     7261 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     7321 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     7381 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     7441 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     7501 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     7561 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     7621 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     7681 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     7741 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     7801 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     7861 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     7921 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     7981 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     8041 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     8101 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     8161 gagcgcaacg caat
//
