LOCUS       sequence-372966-        5061 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vivo visualization of the Golgi apparatus (can be used for 
            colocalization studies).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5061)
  AUTHORS   Bindels DS, Haarbosch L, van Weeren L, Postma M, Wiese KE, Mastop M,
            Aumonier S, Gotthard G, Royant A, Hink MA, Gadella TW Jr
  TITLE     mScarlet: a bright monomeric red fluorescent protein for cellular 
            imaging.
  JOURNAL   Nat Methods. 2016 Nov 21. doi: 10.1038/nmeth.4074.
  PUBMED    27869816
REFERENCE   2  (bases 1 to 5061)
  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: "Nat 
            Methods. 2016 Nov 21. doi: 10.1038/nmeth.4074."
COMMENT     Sequence Label: pmScarlet_Giantin_C1
FEATURES             Location/Qualifiers
     source          1..5061
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        61..165
                     /gene="bla"
                     /label=AmpR promoter
     promoter        167..524
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        167..358
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(190..210)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      375..510
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             559..1353
                     /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             559..1353
                     /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(613..632)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1223..1242
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1541..1560)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1585..1632
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1961..2549
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2450..2469
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2724..3027
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3028..3231
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3182..3202
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3270..3279
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3276..3971
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(3409..3427)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3677..3696
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4512..4633
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4549..4568)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4603..4622
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(join(4640..5061,1..34))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4777..4798)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4989..5008
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg
       61 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
      121 caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga
      181 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
      241 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
      301 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
      361 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
      421 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
      481 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca
      541 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
      601 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
      661 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
      721 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc
      781 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
      841 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
      901 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
      961 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     1021 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     1081 aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     1141 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     1201 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     1261 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     1321 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     1381 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     1441 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     1501 tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa
     1561 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg
     1621 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     1681 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     1741 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct
     1801 caggttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     1861 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     1921 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     1981 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     2041 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     2101 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     2161 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     2221 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     2281 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     2341 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     2401 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     2461 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     2521 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     2581 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     2641 ataaccgtat taccgccatg cattagttat taatagtaat caattacggg gtcattagtt
     2701 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     2761 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     2821 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     2881 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     2941 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     3001 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
     3061 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
     3121 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
     3181 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg
     3241 aaccgtcaga tccgctagcg ctaccggtcg ccaccatggt gagcaagggc gaggcagtga
     3301 tcaaggagtt catgcggttc aaggtgcaca tggagggctc catgaacggc cacgagttcg
     3361 agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg
     3421 tgaccaaggg tggccccctg cccttctcct gggacatcct gtcccctcag ttcatgtacg
     3481 gctccagggc cttcaccaag caccccgccg acatccccga ctactataag cagtccttcc
     3541 ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgcc gtgaccgtga
     3601 cccaggacac ctccctggag gacggcaccc tgatctacaa ggtgaagctc cgcggcacca
     3661 acttccctcc tgacggcccc gtaatgcaga agaagacaat gggctgggaa gcgtccaccg
     3721 agcggttgta ccccgaggac ggcgtgctga agggcgacat taagatggcc ctgcgcctga
     3781 aggacggcgg ccgctacctg gcggacttca agaccaccta caaggccaag aagcccgtgc
     3841 agatgcccgg cgcctacaac gtcgaccgca agttggacat cacctcccac aacgaggact
     3901 acaccgtggt ggaacagtac gaacgctccg agggccgcca ctccaccggc ggcatggacg
     3961 agctgtacaa gtccggactc agatctcgag gagaaccgca gcaaagcttt tctgaagctc
     4021 agcagcagct atgcaacacc agacaggaag tgaatgaatt aaggaagctg ctggaagaag
     4081 aacgagacca aagagtggct gctgagaatg ctctctctgt ggccgaggag cagatcagac
     4141 ggttagagca cagtgaatgg gactcttccc ggactcctat cattggctcc tgtggcactc
     4201 aggagcaggc actgttaata gatcttacaa gcaacagttg tcgaaggacc cggagtggcg
     4261 ttggatggaa gcgagtcctg cgttcactct gtcattcacg gacccgagtg ccacttctag
     4321 cagccatcta ctttctaatg attcatgtcc tgctcattct gtgttttacg ggccatctat
     4381 agggatccac cggatctaga taactgatca taatcagcca taccacattt gtagaggttt
     4441 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     4501 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     4561 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     4621 tcaatgtatc ttaaggcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     4681 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     4741 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     4801 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     4861 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     4921 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     4981 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     5041 acgctgcgcg taaccaccac a
//
