LOCUS       sequence-427396-        7932 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For testing localization of pFAST and iRFP670 proteins to 
            endoplasmic reticulum.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7932)
  TITLE     Tebo lab plasmids 2024
REFERENCE   2  (bases 1 to 7932)
  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"
COMMENT     Sequence Label: ER-FAST-IRES-iRFP670
FEATURES             Location/Qualifiers
     source          1..7932
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             514..1443
                     /codon_start=1
                     /product="near-infrared fluorescent protein with an 
                     emission peak at 670 nm, engineered from a bacterial 
                     phytochrome (Shcherbakova and Verkhusha, 2013)"
                     /label=iRFP670
                     /note="derived from Rhodopseudomonas palustris RpBphP6"
                     /translation="ARKVDLTSCDREPIHIPGSIQPCGCLLACDAQAVRITRITENAGA
                     FFGRETPRVGELLADYFGETEAHALRNALAQSSDPKRPALIFGWRDGLTGRTFDISLHR
                     HDGTSIIEFEPAAAEQADNPLRLTRQIIARTKELKSLEEMAARVPRYLQAMLGYHRVML
                     YRFADDGSGMVIGEAKRSDLESFLGQHFPASLVPQQARLLYLKNAIRVVSDSRGISSRI
                     VPEHDASGAALDLSFAHLRSISPCHLEFLRNMGVSASMSLSIIIDGTLWGLIICHHYEP
                     RAVPMAQRVAAEMFADFLSLHFTAAHHQR"
     CDS             1450..1479
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     polyA_signal    1613..1734
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1650..1669)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1704..1723
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1741..2196)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1878..1899)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2090..2109
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2223..2327
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2329..2686
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2329..2520
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2352..2372)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2537..2672
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2721..3515
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             2721..3515
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2775..2794)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3385..3404
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3703..3722)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3747..3794
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4123..4711
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4612..4631
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4886..5189
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5190..5393
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5344..5364
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     5'UTR           5421..5463
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     CDS             5472..5561
                     /codon_start=1
                     /product="signal sequence from bovine prolactin"
                     /label=prolactin signal sequence
                     /translation="MDSKGSSQKGSRLLLLLVVSNLLLCQGVVS"
     CDS             5574..5948
                     /codon_start=1
                     /product="small protein tag, derived from photoactive 
                     yellow protein, that allows in vivo fluorescent labeling by
                     addition of a cell-permeant fluorogen (Plamont et al., 
                     2016)"
                     /label=Y-FAST
                     /translation="MEHVAFGSEDIENTLANMDDEQLDRLAFGVIQLDGDGNILLYNAA
                     EGDITGRDPKQVIGKNFFKDVAPGTDTPEFYGKFKEGAASGNLNTMFEWTIPTSRGPTK
                     VKVHLKKALSGDRYWVFVKRV"
     CDS             5955..5984
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             6006..6029
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    join(6041..6615,6616..6618,6619..6627)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6208..6225)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     6435..6454
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
ORIGIN
        1 gacaacttta tcatttgtgc tgatcgaaga atagttgatg attgggccaa tgatggatgg
       61 ggcctgaaga aagctgctga tggggctgct gagccaggcg ttgtggggca gatgaacgag
      121 gcagctgaag agcgcccgtg gctgtgggta gtctatattc taactgtagc ccttcctgtg
      181 ttcctggtta tcctcttctg ctgttctgga aagaaacaga ccagtggtat ggagtataag
      241 aaaactgatg cacctcaacc ggatgtgaag gaagaggaag aagagaagga agaggaaaag
      301 gacaagggag atgaggagga ggaaggagaa gagaaacttg aagagaaaca gaaaagtgat
      361 gccgaagaag atggtggcac tgtcagtcaa gaggaggaag acagaaaacc taaagcagag
      421 gaggatgaaa ttttgaacag atcaccaaga aacagaaagc cacgaagaga ggggggtgcg
      481 tcaggcggca gcggatctgg accggtcgct actgcgcgta aggtcgatct cacctcctgc
      541 gatcgcgagc cgatccacat ccccggcagc attcagccgt gcggctgcct gctagcctgc
      601 gacgcgcagg cggtgcggat cacgcgcatt acggaaaatg ccggcgcgtt ctttggacgc
      661 gaaactccgc gggtcggtga gctactcgcc gattacttcg gcgagaccga agcccatgcg
      721 ctgcgcaacg cactggcgca gtcctccgat ccaaagcgac cggcgctgat cttcggttgg
      781 cgcgacggcc tgaccggccg caccttcgac atctcactgc atcgccatga cggtacatcg
      841 atcatcgagt tcgagcctgc ggcggccgaa caggccgaca atccgctgcg gctgacgcgg
      901 cagatcatcg cgcgcaccaa agaactgaag tcgctcgaag agatggccgc acgggtgccg
      961 cgctatctgc aggcgatgct cggctatcac cgcgtgatgt tgtaccgctt cgcggacgac
     1021 ggctccggga tggtgatcgg cgaggcgaag cgcagcgacc tcgagagctt tctcggtcag
     1081 cactttccgg cgtcgctggt cccgcagcag gcgcggctac tgtacttgaa gaacgcgatc
     1141 cgcgtggtct cggattcgcg cggcatcagc agccggatcg tgcccgagca cgacgcctcc
     1201 ggcgccgcgc tcgatctgtc gttcgcgcac ctgcgcagca tctcgccctg ccatctcgaa
     1261 tttctgcgga acatgggcgt cagcgcctcg atgtcgctgt cgatcatcat tgacggcacg
     1321 ctatggggat tgatcatctg tcatcattac gagccgcgtg ccgtgccgat ggcgcagcgc
     1381 gtcgcggccg aaatgttcgc cgacttctta tcgctgcact tcaccgccgc ccaccaccaa
     1441 cgcggatccg aacaaaagct tatttctgaa gaggacttgt gatggatcca ccggatctag
     1501 ataactgatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     1561 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     1621 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     1681 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaacgcgt
     1741 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     1801 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     1861 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     1921 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     1981 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     2041 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     2101 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     2161 cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg
     2221 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     2281 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa
     2341 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     2401 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     2461 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     2521 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     2581 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     2641 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga
     2701 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     2761 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     2821 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     2881 ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg
     2941 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     3001 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     3061 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     3121 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     3181 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     3241 tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     3301 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     3361 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     3421 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     3481 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     3541 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     3601 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     3661 tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac
     3721 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt
     3781 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     3841 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     3901 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     3961 ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     4021 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     4081 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     4141 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     4201 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     4261 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     4321 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     4381 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     4441 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     4501 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     4561 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     4621 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     4681 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     4741 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     4801 ggataaccgt attaccgcca tgcattagtt attaatagta atcaattacg gggtcattag
     4861 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
     4921 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
     4981 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
     5041 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
     5101 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     5161 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     5221 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     5281 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     5341 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag
     5401 tgaaccgtca gatccgctag cttgttcttt ttgcagaagc tcagaataaa cgctcaactt
     5461 tggcagatac catggacagc aaaggttcgt cgcagaaagg gtcccgcctg ctcctgctgc
     5521 tggtggtgtc aaatctactc ttgtgccagg gtgtggtctc ctgcagtcga cttatggagc
     5581 atgttgcctt tggcagtgag gacatcgaga acactctggc caatatggac gacgaacaac
     5641 tggataggtt ggcctttggc gtaattcagc tcgatggtga cgggaatatc ctgctgtaca
     5701 atgctgctga aggggacatc actggcagag atcccaaaca ggtgattggg aagaacttct
     5761 tcaaggatgt tgcacctgga acggatactc ccgagtttta cggcaaattc aaggaaggcg
     5821 cagcgtcagg gaatctgaac accatgttcg aatggacgat accgacaagc aggggaccaa
     5881 ccaaggtcaa ggtgcacttg aagaaagccc tttccggtga cagatattgg gtctttgtga
     5941 aacgggtggg atccgaacaa aagcttattt ctgaagagga cttgaaagat gagctctaac
     6001 tcgaggacta caaggacgac gacgacaagc ccgggatccg cccctctccc tccccccccc
     6061 ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc tatatgttat
     6121 tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc cctgtcttct
     6181 tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg
     6241 tcgtgaagga agcagttcct ctggaagctt cttgaagaca aacaacgtct gtagcgaccc
     6301 tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa aagccacgtg
     6361 tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt tggatagttg
     6421 tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag gatgcccaga
     6481 aggtacccca ttgtatggga tctgatctgg ggcctcggta cacatgcttt acatgtgttt
     6541 agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa
     6601 aaacacgatg ataatatggc cacaaccatg gaagggaagt ggttgctgtg tatgttactg
     6661 gtgcttggaa ctgctattgt tgaggctcat gatggacatg atgatgatgt gattgatatt
     6721 gaggatgacc ttgacgatgt cattgaagag gtagaagact caaaaccaga taccactgct
     6781 cctccttcat ctcccaaggt tacttacaaa gctccagttc caacagggga agtatatttt
     6841 gctgattctt ttgacagagg aactctgtca gggtggattt tatccaaagc caagaaagac
     6901 gataccgatg atgaaattgc caaatatgat ggaaagtggg aggtagagga aatgaaggag
     6961 tcaaagcttc caggtgataa aggacttgtg ttgatgtctc gggccaagca tcatgccatc
     7021 tctgctaaac tgaacaagcc cttcctgttt gacaccaagc ctctcattgt tcagtatgag
     7081 gttaatttcc aaaatggaat agaatgtggt ggtgcctatg tgaaactgct ttctaaaaca
     7141 ccagaactca acctggatca gttccatgac aagacccctt atacgattat gtttggtcca
     7201 gataaatgtg gagaggacta taaactgcac ttcatcttcc gacacaaaaa ccccaaaacg
     7261 ggtatctatg aagaaaaaca tgctaagagg ccagatgcag atctgaagac ctattttact
     7321 gataagaaaa cacatcttta cacactaatc ttgaatccag ataatagttt tgaaatactg
     7381 gttgaccaat ctgtggtgaa tagtggaaat ctgctcaatg acatgactcc tcctgtaaat
     7441 ccttcacgtg aaattgagga cccagaagac cggaagcccg aggattggga tgaaagacca
     7501 aaaatcccag atccagaagc tgtcaagcca gatgactggg atgaagatgc ccctgctaag
     7561 attccagatg aagaggccac aaaacccgaa ggctggttag atgatgagcc tgagtacgta
     7621 cctgatccag acgcagagaa acctgaggat tgggatgaag acatggatgg agaatgggag
     7681 gctcctcaga ttgccaaccc tagatgtgag tcagctcctg gatgtggtgt ctggcagcga
     7741 cctgtgattg acaaccccaa ttataaaggc aaatggaagc ctcctatgat tgacaatccc
     7801 agttaccagg gaatctggaa acccaggaaa ataccaaatc cagatttctt tgaagatctg
     7861 gaacctttca gaatgactcc ttttagtgct attggtttgg agctgtggtc catgacctct
     7921 gacatttttt tt
//
