LOCUS       sequence-458169-        5786 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing fluorescent HaloTag7 fusion protein in mammalian cells 
            with localization in the Golgi apparatus..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5786)
  AUTHORS   Miro-Vinyals C, Emmert S, Grammbitter G, Rivera-Fuentes P
  TITLE     Characterization of the Glutathione Redox State in the Golgi 
            Apparatus
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5786)
  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: 
            "Unpublished"
COMMENT     Sequence Label: Galt1-HT7-mGold
FEATURES             Location/Qualifiers
     source          1..5786
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1061..1264)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1090..1110)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1265..1568
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(1743..2331)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1823..1842)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2660..2707
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     2732..2751
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(2939..3733)
                     /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             complement(2939..3733)
                     /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(3050..3069)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3660..3679
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(3768..4125)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3782..3917
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        3934..4125
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4082..4102
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(4127..4231)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4258..4713
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4345..4364)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4555..4576
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4720..4841
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4731..4750)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4785..4804
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(4964..5677,5678..5680))
                     /codon_start=1
                     /product="photostable derivative of mVenus (Lee et al., 
                     2020)"
                     /label=mGold
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTSLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(5009..5030)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     5358..5377
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5617..5638
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(join(5681..5786,1..779))
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
ORIGIN
        1 cagttaggca ggcttttggc caggcgagcg gcttcggccg gtgggatcag aacgcctggg
       61 gtgccccaga acagcagctt cgggacaggg gactggtgca gccagtccat gtattcttcg
      121 accagcgcga cgatgttcgc tggctcaccg gcgattggca gctcgtttgg gaagcgccac
      181 agtggctcgc ggtcaacagg attcaggaac ggctcgcggt aatggtccat ctcgacttca
      241 gtcagcgggc ggacgacacc catcggcagc gtaccctcga taaaaacgtt ctgatcgatg
      301 atcagcttgc ggccgacgtc ggtggtgcgg aaggcctgga aggtctcgcg ggcaaattct
      361 ggccattcgt cccaggtcgg gatagggcgg atgaactcca taaatgcaat acctttgacg
      421 cgctctggat tgcgcttggc ccagtggaaa cccagagcgg agccccagtc gtgaatgacc
      481 aggacgacct cttccagacc cagggcttcg atgaaggcat ccatgaagcg gacgtggtcg
      541 tcgaagaaat aacccaggtc tggtttgtcg gatttgccca taccgatcag gtctggagca
      601 atgcagcgat gggtcggtgc aacatgcggg atgatgttgc gccacacgta ggaggaggtc
      661 gggttaccgt gcaggaacag cacaggggtg ccatcgcgcg gaccaacatc gacgtagtgc
      721 atgcgctcgc ccaggacttc cacataatgg gggtcgaatg gaaagccagt accgatttcg
      781 gtggcgaccg gtggatcccg ggcccgcggt accgtcgact gcagcggtgt ggagactccg
      841 accagttggg gcaggcggct caggtcgcgg ccagccaggt agtaaacgag ggtgacgcca
      901 aggtgcagag cgcagacggc cacgagcagg cggcaggccc gctgtaggga cgcgcctggc
      961 atcgcggcgc tgccgctcag gagcggctcc cgaagcctca tggtggcgag ctcgagatct
     1021 gagtccggta gcgctagcgg atctgacggt tcactaaacc agctctgctt atatagacct
     1081 cccaccgtac acgcctaccg cccatttgcg tcaatggggc ggagttgtta cgacattttg
     1141 gaaagtcccg ttgattttgg tgccaaaaca aactcccatt gacgtcaatg gggtggagac
     1201 ttggaaatcc ccgtgagtca aaccgctatc cacgcccatt gatgtactgc caaaaccgca
     1261 tcaccatggt aatagcgatg actaatacgt agatgtactg ccaagtagga aagtcccata
     1321 aggtcatgta ctgggcataa tgccaggcgg gccatttacc gtcattgacg tcaatagggg
     1381 gcgtacttgg catatgatac acttgatgta ctgccaagtg ggcagtttac cgtaaatact
     1441 ccacccattg acgtcaatgg aaagtcccta ttggcgttac tatgggaaca tacgtcatta
     1501 ttgacgtcaa tgggcggggg tcgttgggcg gtcagccagg cgggccattt accgtaagtt
     1561 atgtaacgcg gaactccata tatgggctat gaactaatga ccccgtaatt gattactatt
     1621 aataactaat gcatggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     1681 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     1741 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     1801 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     1861 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     1921 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     1981 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     2041 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     2101 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     2161 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     2221 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     2281 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     2341 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     2401 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     2461 aaatcaatct aaagtatata tgagtaacct gaggctatgg cagggcctgc cgccccgacg
     2521 ttggctgcga gccctgggcc ttcacccgaa cttggggggt ggggtgggga aaaggaagaa
     2581 acgcgggcgt attggcccca atggggtctc ggtggggtat cgacagagtg ccagccctgg
     2641 gaccgaaccc cgcgtttatg aacaaacgac ccaacaccgt gcgttttatt ctgtcttttt
     2701 attgccgtca tagcgcgggt tccttccggt attgtctcct tccgtgtttc agttagcctc
     2761 cccctagggt gggcgaagaa ctccagcatg agatccccgc gctggaggat catccagccg
     2821 gcgtcccgga aaacgattcc gaagcccaac ctttcataga aggcggcggt ggaatcgaaa
     2881 tctcgtgatg gcaggttggg cgtcgcttgg tcggtcattt cgaaccccag agtcccgctc
     2941 agaagaactc gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga gcggcgatac
     3001 cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag ctcttcagca atatcacggg
     3061 tagccaacgc tatgtcctga tagcggtccg ccacacccag ccggccacag tcgatgaatc
     3121 cagaaaagcg gccattttcc accatgatat tcggcaagca ggcatcgcca tgggtcacga
     3181 cgagatcctc gccgtcgggc atgctcgcct tgagcctggc gaacagttcg gctggcgcga
     3241 gcccctgatg ctcttcgtcc agatcatcct gatcgacaag accggcttcc atccgagtac
     3301 gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg
     3361 tatgcagccg ccgcattgca tcagccataa tggatacttt ctcggcagga gcaaggtgag
     3421 atgacaggag atcctgcccc ggcacttcgc ccaatagcag ccagtccctt cccgcttcag
     3481 tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg
     3541 ctgcctcgtc ttgcagttca ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg
     3601 ggcgcccctg cgctgacagc cggaacacgg cggcatcaga gcagccgatt gtctgttgtg
     3661 cccagtcata gccgaatagc ctctccaccc aagcggccgg agaacctgcg tgcaatccat
     3721 cttgttcaat catgcgaaac gatcctcatc ctgtctcttg atcgatcttt gcaaaagcct
     3781 aggcctccaa aaaagcctcc tcactacttc tggaatagct cagaggccga ggcggcctcg
     3841 gcctctgcat aaataaaaaa aattagtcag ccatggggcg gagaatgggc ggaactgggc
     3901 ggagttaggg gcgggatggg cggagttagg ggcgggacta tggttgctga ctaattgaga
     3961 tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca cctggttgct
     4021 gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca
     4081 caccctaact gacacacatt ccacagctgg ttctttccgc ctcaggactc ttcctttttc
     4141 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     4201 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     4261 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     4321 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     4381 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     4441 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     4501 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     4561 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     4621 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     4681 cgaattttaa caaaatatta acgcttacaa tttacgcctt aagatacatt gatgagtttg
     4741 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     4801 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     4861 attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct
     4921 acaaatgtgg tatggctgat tatgatctag agtcgcggcc gctttacttg tacagctcgt
     4981 ccatgccgag agtgatcccg gcggcggtca cgaactccag caggaccatg tgatcgcgct
     5041 tctcgttggg gtctttgctc agcttggact ggtagctcag gtagtggttg tcgggcagca
     5101 gcacggggcc gtcgccgatg ggggtgttct gctggtagtg gtcggcgagc tgcacgccgc
     5161 cgtcctcgat gttgtggcgg atcttgaagt tggccttgat gccgttcttc tgcttgtcgg
     5221 cggtgatata gacgttgtgg ctgttgtagt tgtactccag cttgtgcccc aggatgttgc
     5281 cgtcctcctt gaagtcgatg cccttcagct cgatgcggtt caccagggtg tcgccctcga
     5341 acttcacctc ggcgcgggtc ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct
     5401 ggacgtagcc ttcgggcatg gcggacttga agaagtcgtg ctgcttcatg tggtcggggt
     5461 agcgggcgaa gcactgcagg ccgtagccca ggctggtcac gagggtgggc cagggcacgg
     5521 gcagcttgcc ggtggtgcag atgaacttca gggtcagctt gccgtaggtg gcatcgccct
     5581 cgccctcgcc ggacacgctg aacttgtggc cgtttacgtc gccgtccagc tcgaccagga
     5641 tgggcaccac cccggtgaac agctcctcgc ccttgctcac gccggaaatc tcgagcgtgg
     5701 acagccagcg cgcgatctcg ctgccgatca ggtccgggtt gtcttcttgc agcagattca
     5761 gacccgggcc gatgtccaca gccttg
//
