LOCUS       sequence-427414-        6484 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For testing localization of pFAST and iRFP670 proteins to Golgi 
            complex.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6484)
  TITLE     Tebo lab plasmids 2024
REFERENCE   2  (bases 1 to 6484)
  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: Golgi-FAST-IRES-iRFP670
FEATURES             Location/Qualifiers
     source          1..6484
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             122..916
                     /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             122..916
                     /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(176..195)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     786..805
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1104..1123)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1148..1195
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1524..2112
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2013..2032
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2287..2590
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2591..2794
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2745..2765
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3028..3037
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3034..3408
                     /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             3415..3444
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3454..3477
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    join(3489..4063,4064..4066,4067..4075)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3656..3673)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3883..3902
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4571..5500
                     /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             5507..5536
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     polyA_signal    5570..5691
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5607..5626)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5661..5680
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5698..6153)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5835..5856)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6047..6066
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6180..6284
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6309..6329)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(6353..6484,1..79)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        join(6375..6484,1..87)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      join(6422..6484,1..73)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
       61 ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg
      121 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
      181 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
      241 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
      301 gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
      361 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
      421 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
      481 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
      541 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
      601 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga
      661 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
      721 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
      781 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
      841 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
      901 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
      961 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     1021 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     1081 gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     1141 gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa
     1201 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
     1261 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
     1321 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata
     1381 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     1441 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     1501 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     1561 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     1621 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     1681 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     1741 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     1801 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     1861 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     1921 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     1981 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     2041 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     2101 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     2161 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     2221 atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
     2281 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     2341 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     2401 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     2461 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     2521 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     2581 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
     2641 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
     2701 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
     2761 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta
     2821 gcgctaccgg actcagatct cgagctcgcc accatgattc acaccaacct gaagaaaaag
     2881 ttcagctgct gcgtcctggt ctttcttctg tttgcagtca tctgtgtgtg gaaggaaaag
     2941 aagaaaggga gttactatga ttcctttaaa ttgcaaacca aggaattcct gcagtcgacg
     3001 gtaccgcggg cccgggatcc accggtcgcc accatggagc atgttgcctt tggcagtgag
     3061 gacatcgaga acactctggc caatatggac gacgaacaac tggataggtt ggcctttggc
     3121 gtaattcagc tcgatggtga cgggaatatc ctgctgtaca atgctgctga aggggacatc
     3181 actggcagag atcccaaaca ggtgattggg aagaacttct tcaaggatgt tgcacctgga
     3241 acggatactc ccgagtttta cggcaaattc aaggaaggcg cagcgtcagg gaatctgaac
     3301 accatgttcg aatggacgat accgacaagc aggggaccaa ccaaggtcaa ggtgcacttg
     3361 aagaaagccc tttccggtga cagatattgg gtctttgtga aacgggtggg atccgaacaa
     3421 aagcttattt ctgaagagga cttgtaactc gaggactaca aggacgacga cgacaagccc
     3481 gggatccgcc cctctccctc ccccccccct aacgttactg gccgaagccg cttggaataa
     3541 ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg
     3601 agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc
     3661 gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct
     3721 tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac
     3781 aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc
     3841 cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta
     3901 ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg
     3961 cctcggtaca catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg
     4021 aaccacgggg acgtggtttt cctttgaaaa acacgatgat aatatggcca caaccatggc
     4081 gggtcagttc cgcagctacg tgtgggaccc gctgctgatc ctgtcgcaga tcgtcctcat
     4141 gcagaccgtg tattacggct cgctgggcct gtggctggcg ctggtggacg ggctagtgcg
     4201 aagcagcccc tcgctggacc agatgttcga cgccgagatc ctgggctttt ccacccctcc
     4261 aggccggctc tccatgatgt ccttcatcct caacgccctc acctgtgccc tgggcttgct
     4321 gtacttcatc cggcgaggaa agcagtgtct ggatttcact gtcactgtcc atttctttca
     4381 cctcctgggc tgctggttct acagctcccg tttcccctcg gcgctgacct ggtggctggt
     4441 ccaagccgtg tgcattgcac tcatggctgt catcggggag tacctgtgca tgcggacgga
     4501 gctcaaggag atacccctca actcagcccc taaatccaat gtccgggatc caccggtcgc
     4561 caccgctact gcgcgtaagg tcgatctcac ctcctgcgat cgcgagccga tccacatccc
     4621 cggcagcatt cagccgtgcg gctgcctgct agcctgcgac gcgcaggcgg tgcggatcac
     4681 gcgcattacg gaaaatgccg gcgcgttctt tggacgcgaa actccgcggg tcggtgagct
     4741 actcgccgat tacttcggcg agaccgaagc ccatgcgctg cgcaacgcac tggcgcagtc
     4801 ctccgatcca aagcgaccgg cgctgatctt cggttggcgc gacggcctga ccggccgcac
     4861 cttcgacatc tcactgcatc gccatgacgg tacatcgatc atcgagttcg agcctgcggc
     4921 ggccgaacag gccgacaatc cgctgcggct gacgcggcag atcatcgcgc gcaccaaaga
     4981 actgaagtcg ctcgaagaga tggccgcacg ggtgccgcgc tatctgcagg cgatgctcgg
     5041 ctatcaccgc gtgatgttgt accgcttcgc ggacgacggc tccgggatgg tgatcggcga
     5101 ggcgaagcgc agcgacctcg agagctttct cggtcagcac tttccggcgt cgctggtccc
     5161 gcagcaggcg cggctactgt acttgaagaa cgcgatccgc gtggtctcgg attcgcgcgg
     5221 catcagcagc cggatcgtgc ccgagcacga cgcctccggc gccgcgctcg atctgtcgtt
     5281 cgcgcacctg cgcagcatct cgccctgcca tctcgaattt ctgcggaaca tgggcgtcag
     5341 cgcctcgatg tcgctgtcga tcatcattga cggcacgcta tggggattga tcatctgtca
     5401 tcattacgag ccgcgtgccg tgccgatggc gcagcgcgtc gcggccgaaa tgttcgccga
     5461 cttcttatcg ctgcacttca ccgccgccca ccaccaacgc ggatccgaac aaaagcttat
     5521 ttctgaagag gacttgtaat taattaagca agggcgaatt gttgttgtta acttgtttat
     5581 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     5641 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa
     5701 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     5761 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     5821 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     5881 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     5941 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     6001 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     6061 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     6121 ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc
     6181 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     6241 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa
     6301 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     6361 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     6421 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     6481 tttt
//
