LOCUS       sequence-339210-        7251 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FKBP anchor for retaining DHFR-fused cargoes in the ER in the 
            presence of zapalog. Encodes ER retrieval motif (KDEL) at C-terminus
            of spaghetti monster FP (myc, non-fluorescent). IRES following ORF..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7251)
  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 7251)
  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: 2xFKBP-smFP(myc)-KDEL
FEATURES             Location/Qualifiers
     source          1..7251
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..638
                     /label=chicken beta-actin promoter
                     /note="contains a 21-bp deletion relative to the standard 
                     chicken beta-actin promoter"
     intron          639..1655
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1579..1601
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1663..1682
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     sig_peptide     1716..1775
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             1779..1805
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1812..2132
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             2160..2480
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             2517..2546
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2550..2579
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2583..2612
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3144..3173
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3180..3209
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3222..3251
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3258..3287
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    3534..4107
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3701..3718)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3928..3947
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     complement(4239..4258)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4304..4359
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4358..4377)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4750..4766)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4750..4766)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4763..4785)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4774..4790
                     /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(4798..4804,4805..4822,4823..4828))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4843..4864
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4923..5118
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4969..5104
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5124..5258
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5161..5180)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5215..5234
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5326..5343)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5497..6085)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5577..5596)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6256..7047,7048..7116))
                     /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     6879..6898
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7117..7221)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
      421 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
      481 tgggggcggg gggggggggg gggcggggcg aggggcgggg cggggcgagg cggagaggtg
      541 cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc
      601 ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg agtcgctgcg cgctgccttc
      661 gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt
      721 tactcccaca ggtgagcggg cgggacggcc cttctcctcc gggctgtaat tagcgcttgg
      781 tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag ccttgagggg ctccgggagg
      841 gccctttgtg cggggggagc ggctcggggg gtgcgtgcgt gtgtgtgtgc gtggggagcg
      901 ccgcgtgcgg ctccgcgctg cccggcggct gtgagcgctg cgggcgcggc gcggggcttt
      961 gtgcgctccg cagtgtgcgc gaggggagcg cggccggggg cggtgccccg cggtgcgggg
     1021 ggggctgcga ggggaacaaa ggctgcgtgc ggggtgtgtg cgtggggggg tgagcagggg
     1081 gtgtgggcgc gtcggtcggg ctgcaacccc ccctgcaccc ccctccccga gttgctgagc
     1141 acggcccggc ttcgggtgcg gggctccgta cggggcgtgg cgcggggctc gccgtgccgg
     1201 gcggggggtg gcggcaggtg ggggtgccgg gcggggcggg gccgcctcgg gccggggagg
     1261 gctcggggga ggggcgcggc ggcccccgga gcgccggcgg ctgtcgaggc gcggcgagcc
     1321 gcagccattg ccttttatgg taatcgtgcg agagggcgca gggacttcct ttgtcccaaa
     1381 tctgtgcgga gccgaaatct gggaggcgcc gccgcacccc ctctagcggg cgcggggcga
     1441 agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg ccttcgtgcg tcgccgcgcc
     1501 gccgtcccct tctccctctc cagcctcggg gctgtccgcg gggggacggc tgccttcggg
     1561 ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac cggcggctct agagcctctg
     1621 ctaaccatgt tcatgccttc ttctttttcc tacagctcct gggcaacgtg ctggttattg
     1681 tgctgtctca tcattttggc aaagaattgc ccaccatgga gacagacaca ctcctgctat
     1741 gggtactgct gctctgggtt ccaggttcca ctggtgacta tccatatgat gttccagatt
     1801 atgctctcga gggagtccag gtagagacga tttcacccgg cgacggccgc acattcccga
     1861 agcgaggcca gacatgtgtg gtgcattata ccgggatgct tgaggatgga aagaagttcg
     1921 actcaagtag ggatcggaac aaaccgttca agttcatgtt gggaaagcaa gaagtaatac
     1981 ggggttggga ggaaggtgtt gcccaaatga gtgttggtca gagagctaag ctgacgatca
     2041 gtcccgacta tgcctatggt gcaacaggac atccaggaat aatacctccg cacgccactt
     2101 tggtattcga tgtcgaactc ttgaagctgg aaagtgctgg tagtgctggt agtgctggtg
     2161 gagtgcaggt ggaaaccatc tccccaggag acgggcgcac cttccccaag cgcggccaga
     2221 cctgcgtggt gcactacacc gggatgcttg aagatggaaa gaaatttgat tcctcccggg
     2281 acagaaacaa gccctttaag tttatgctag gcaagcagga ggtgatccga ggctgggaag
     2341 aaggggttgc ccagatgagt gtgggtcaga gagccaaact gactatatct ccagattatg
     2401 cctatggtgc cactgggcac ccaggcatca tcccaccaca tgccactctc gtcttcgatg
     2461 tggagcttct aaaactggaa ggtggcggcg gaagcggtgg cgggggtgga ccggtcgagc
     2521 aaaaactgat atcggaggaa gacttggcag agcagaaact gatttcggaa gaggatcttg
     2581 ccgaacaaaa actgatcagt gaggaagacc ttgccggagt gtcgaagggt gaagagctgt
     2641 tcaccggcgt ggtgcccatc ctcgtcgagc tggacggaga tgtcaatgga cataaattct
     2701 ccgtacgggg agagggcgag ggcgacgcaa ccaacggtaa actgaccctg aagttcattt
     2761 gcaccacggg taaactgccc gtaccctggc ccaccctggt cacaaccttg ggtggcggag
     2821 ttcagtgttt ttcgcggtat ccagatcaca tgaagcagca tgactttttc aagagcgcta
     2881 tgcccgaggg ttacgttcaa gagaggacca tcagtttcaa ggacgacggc acctacaaga
     2941 cacgggctga ggtgaaattc gagggtgata cactcgtgaa tcgtatcgag ctgaagggca
     3001 tagattttaa ggaggatggc aatatcctgg gccacaagtt ggagtataac ttcaacagtc
     3061 acaatgtcta tataactgcc gataagcaga agaatggcat taaagccaat ttcaaaattc
     3121 gtcacaatgt cgaaggcggc gcagaacaga agctcatcag tgaagaggat ctggccgcag
     3181 agcagaaact cattagcgag gaggacttgg gcggaggcgg cgagcaaaaa ctgatatcgg
     3241 aggaagacct ggcggccgag caaaagctca tctcggagga ggatctcgcg ggaggagatg
     3301 gaagtgtcca gttggcggat cactaccagc agaatacgcc aattggtgac ggccccgtcc
     3361 ttctgccgga taaccactac ctctccaccc agtcggtgct gagcaaggac ccgaatgaaa
     3421 aacgtgacca catggttctt cttgagttcg tcacggctgc gggcatcacg ttgggcatgg
     3481 acgaactgta caagaaggat gagctgtaaa cggtaccgcg ggcccgggat ccgcccctct
     3541 ccctcccccc cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt
     3601 gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct
     3661 ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa
     3721 ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg
     3781 tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc
     3841 caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg
     3901 agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg
     3961 aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtacacatgc
     4021 tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg
     4081 gttttccttt gaaaaacacg atgataatga attctgcagt cgacggtacc gcgggcccta
     4141 caagtaaagc ggccgcaatt cactcctcag gtgcaggctg cctatcagaa ggtggtggct
     4201 ggtgtggcca atgccctggc tcacaaatac cactgagatc tttttccctc tgccaaaaat
     4261 tatggggaca tcatgaagcc ccttgagcat ctgacttctg gctaataaag gaaatttatt
     4321 ttcattgcaa tagtgtgttg gaattttttg tgtctctcac tcggaaggac atatgggagg
     4381 gcaaatcatt taaaacatca gaatgagtat ttggtttaga gtttggcaac atatgcccat
     4441 atgctggctg ccatgaacaa aggttggcta taaagaggtc atcagtatat gaaacagccc
     4501 cctgctgtcc attccttatt ccatagaaaa gccttgactt gaggttagat tttttttata
     4561 ttttgttttg tgttattttt ttctttaaca tccctaaaat tttccttaca tgttttacta
     4621 gccagatttt tcctcctctc ctgactactc ccagtcatag ctgtccctct tctcttatgg
     4681 agatccctcg acctgcaccg tcgaccagct ggtcgacggt gcaccgtcga ccagcttggc
     4741 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     4801 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     4861 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagcggat
     4921 ccgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     4981 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     5041 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     5101 ggcctaggct tttgcaaaaa gctaacttgt ttattgcagc ttataatggt tacaaataaa
     5161 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     5221 tgtccaaact catcaatgta tcttatcatg tctggatccg ctgcattaat gaatcggcca
     5281 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     5341 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     5401 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     5461 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     5521 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     5581 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     5641 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     5701 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     5761 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     5821 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     5881 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     5941 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     6001 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     6061 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     6121 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     6181 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     6241 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     6301 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     6361 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     6421 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     6481 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     6541 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     6601 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     6661 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     6721 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     6781 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     6841 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     6901 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6961 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     7021 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     7081 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     7141 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     7201 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgggt c
//
