LOCUS       sequence-427413-        4464 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For testing localization of pFAST protein to Golgi complex.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4464)
  TITLE     Tebo lab plasmids 2024
REFERENCE   2  (bases 1 to 4464)
  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
FEATURES             Location/Qualifiers
     source          1..4464
                     /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"
     polyA_signal    3478..3599
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3515..3534)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3569..3588
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3606..4061)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3743..3764)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3955..3974
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4088..4192
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(4194..4464,1..87)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4194..4385
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4217..4237)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      join(4402..4464,1..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 cttgtaatta attaagcaag ggcgaattgt tgttgttaac
     3481 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     3541 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa
     3601 ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     3661 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     3721 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     3781 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     3841 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     3901 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     3961 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     4021 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg
     4081 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     4141 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc
     4201 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     4261 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     4321 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     4381 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     4441 ccccatggct gactaatttt tttt
//
