LOCUS       sequence-458168-        4577 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing sCGrx1p protein fused to HA tag 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 4577)
  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  JOURNAL   Unpublished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 4577)
  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            "Unpublished"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-sCGrx1p-HA
FEATURES             Location/Qualifiers
     source          1..4577
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(136..162)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      165..174
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(798..1001)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(827..847)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1002..1305
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(1480..2068)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1560..1579)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2397..2444
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     2469..2488
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(2676..3470)
                     /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(2676..3470)
                     /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(2787..2806)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3397..3416
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(3505..3862)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3519..3654
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        3671..3862
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     3819..3839
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(3864..3968)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3995..4450
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4082..4101)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4292..4313
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    join(4457..4577,1)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4468..4487)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4522..4541
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
       61 ttaaagcaag taaaacctct acaaatgtgg tatggctgat tatgatctag agtcgcggcc
      121 gctttaggta ccttctgcgt agtcaggcac atcatacgga tacaccatgg tggcgacatt
      181 tgcaagaata ggttctaaca attcctccaa ttcaccagtc tccctcaatt cctgcaagtc
      241 gtcgttgcct ccaatatgtt taccattaat atagatgttt ggcacggttc tttggccatt
      301 aatctcatat aacgcagcct gaatgtctgc gccttccttc atgtcattca attgcaaaac
      361 cagaactttg gacctgggaa cctttaactt ttcaaaaagc gtgtttaaag ctgcatgaga
      421 gtatggacag tacgttttgg atgcgacgaa gatctcgttt tctgcaataa ggtccttgac
      481 gtgcttgata gtttcttgag ataccatggt accttcggtg gcgaccggtg gatcccgggc
      541 ccgcggtacc gtcgactgca gcggtgtgga gactccgacc agttggggca ggcggctcag
      601 gtcgcggcca gccaggtagt aaacgagggt gacgccaagg tgcagagcgc agacggccac
      661 gagcaggcgg caggcccgct gtagggacgc gcctggcatc gcggcgctgc cgctcaggag
      721 cggctcccga agcctcatgg tggcgagctc gagatctgag tccggtagcg ctagcggatc
      781 tgacggttca ctaaaccagc tctgcttata tagacctccc accgtacacg cctaccgccc
      841 atttgcgtca atggggcgga gttgttacga cattttggaa agtcccgttg attttggtgc
      901 caaaacaaac tcccattgac gtcaatgggg tggagacttg gaaatccccg tgagtcaaac
      961 cgctatccac gcccattgat gtactgccaa aaccgcatca ccatggtaat agcgatgact
     1021 aatacgtaga tgtactgcca agtaggaaag tcccataagg tcatgtactg ggcataatgc
     1081 caggcgggcc atttaccgtc attgacgtca atagggggcg tacttggcat atgatacact
     1141 tgatgtactg ccaagtgggc agtttaccgt aaatactcca cccattgacg tcaatggaaa
     1201 gtccctattg gcgttactat gggaacatac gtcattattg acgtcaatgg gcgggggtcg
     1261 ttgggcggtc agccaggcgg gccatttacc gtaagttatg taacgcggaa ctccatatat
     1321 gggctatgaa ctaatgaccc cgtaattgat tactattaat aactaatgca tggcggtaat
     1381 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     1441 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     1501 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     1561 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     1621 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     1681 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     1741 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     1801 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     1861 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     1921 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     1981 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     2041 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     2101 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     2161 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     2221 gtaacctgag gctatggcag ggcctgccgc cccgacgttg gctgcgagcc ctgggccttc
     2281 acccgaactt ggggggtggg gtggggaaaa ggaagaaacg cgggcgtatt ggccccaatg
     2341 gggtctcggt ggggtatcga cagagtgcca gccctgggac cgaaccccgc gtttatgaac
     2401 aaacgaccca acaccgtgcg ttttattctg tctttttatt gccgtcatag cgcgggttcc
     2461 ttccggtatt gtctccttcc gtgtttcagt tagcctcccc ctagggtggg cgaagaactc
     2521 cagcatgaga tccccgcgct ggaggatcat ccagccggcg tcccggaaaa cgattccgaa
     2581 gcccaacctt tcatagaagg cggcggtgga atcgaaatct cgtgatggca ggttgggcgt
     2641 cgcttggtcg gtcatttcga accccagagt cccgctcaga agaactcgtc aagaaggcga
     2701 tagaaggcga tgcgctgcga atcgggagcg gcgataccgt aaagcacgag gaagcggtca
     2761 gcccattcgc cgccaagctc ttcagcaata tcacgggtag ccaacgctat gtcctgatag
     2821 cggtccgcca cacccagccg gccacagtcg atgaatccag aaaagcggcc attttccacc
     2881 atgatattcg gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc gtcgggcatg
     2941 ctcgccttga gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga
     3001 tcatcctgat cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc
     3061 gcttggtggt cgaatgggca ggtagccgga tcaagcgtat gcagccgccg cattgcatca
     3121 gccatgatgg atactttctc ggcaggagca aggtgagatg acaggagatc ctgccccggc
     3181 acttcgccca atagcagcca gtcccttccc gcttcagtga caacgtcgag cacagctgcg
     3241 caaggaacgc ccgtcgtggc cagccacgat agccgcgctg cctcgtcttg cagttcattc
     3301 agggcaccgg acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg
     3361 aacacggcgg catcagagca gccgattgtc tgttgtgccc agtcatagcc gaatagcctc
     3421 tccacccaag cggccggaga acctgcgtgc aatccatctt gttcaatcat gcgaaacgat
     3481 cctcatcctg tctcttgatc gatctttgca aaagcctagg cctccaaaaa agcctcctca
     3541 ctacttctgg aatagctcag aggccgaggc ggcctcggcc tctgcataaa taaaaaaaat
     3601 tagtcagcca tggggcggag aatgggcgga actgggcgga gttaggggcg ggatgggcgg
     3661 agttaggggc gggactatgg ttgctgacta attgagatgc atgctttgca tacttctgcc
     3721 tgctggggag cctggggact ttccacacct ggttgctgac taattgagat gcatgctttg
     3781 catacttctg cctgctgggg agcctgggga ctttccacac cctaactgac acacattcca
     3841 cagctggttc tttccgcctc aggactcttc ctttttcaat attattgaag catttatcag
     3901 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     3961 gttccgcgca catttccccg aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc
     4021 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     4081 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     4141 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     4201 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     4261 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     4321 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     4381 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     4441 cttacaattt acgccttaag atacattgat gagtttggac aaaccacaac tagaatgcag
     4501 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     4561 agctgcaata aacaagt
//
