LOCUS       sequence-275933-        7810 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Golgi marker GalnacT2-msGFP2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7810)
  AUTHORS   Casler JC, Zajac AL, Valbuena FM, Sparvoli D, Jeyifous O, Turkewitz 
            AP, Horne-Badovinac S, Green WN, Glick BS
  TITLE     ESCargo: a regulatable fluorescent secretory cargo for diverse model
            organisms.
  JOURNAL   Mol Biol Cell. 2020 Oct 28:mbcE20090591. doi: 
            10.1091/mbc.E20-09-0591.
  PUBMED    33112725
REFERENCE   2  (bases 1 to 7810)
  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: "Mol Biol 
            Cell. 2020 Oct 28:mbcE20090591. doi: 10.1091/mbc.E20-09-0591."
COMMENT     Sequence Label: pEF-GalNAc-T2-msGFP2
FEATURES             Location/Qualifiers
     source          1..7810
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        6..1168
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          237..1159
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1116..1136
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             2919..3626
                     /codon_start=1
                     /product="photostable monomeric superfolder GFP (Valbuena 
                     et al., 2020)"
                     /label=msGFP2
                     /translation="MDSTESLFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTITFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITGGSGGS"
     primer_bind     complement(2961..2982)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     3569..3590
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3650..3667)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3656..3880
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3926..4354
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4013..4032)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4223..4244
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4363..4383)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4368..4697
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4548..4683
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4610..4629
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4723..5517
                     /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="MVEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4777..4796)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5387..5406
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5691..5812
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5728..5747)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5782..5801
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5845..5862)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6016..6604)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6096..6115)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6775..7566,7567..7635))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7398..7417
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7636..7740)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cgtgaggctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt
       61 tggggggagg ggtcggcaat tgaaccggtg cctagagaag gtggcgcggg gtaaactggg
      121 aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa
      181 gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacaggtaa
      241 gtgccgtgtg tggttcccgc gggcctggcc tctttacggg ttatggccct tgcgtgcctt
      301 gaattacttc cacctggctc cagtacgtga ttcttgatcc cgagctggag ccaggggcgg
      361 gccttgcgct ttaggagccc cttcgcctcg tgcttgagtt gaggcctggc ctgggcgctg
      421 gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt tcgataagtc
      481 tctagccatt taaaattttt gatgacctgc tgcgacgctt tttttctggc aagatagtct
      541 tgtaaatgcg ggccaggatc tgcacactgg tatttcggtt tttggggccg cgggcggcga
      601 cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc
      661 gagaatcgga cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg gcctcgcgcc
      721 gccgtgtatc gccccgccct gggcggcaag gctggcccgg tcggcaccag ttgcgtgagc
      781 ggaaagatgg ccgcttcccg gccctgctcc agggggctca aaatggagga cgcggcgctc
      841 gggagagcgg gcgggtgagt cacccacaca aaggaaaggg gcctttccgt cctcagccgt
      901 cgcttcatgt gactccacgg agtaccgggc gccgtccagg cacctcgatt agttctggag
      961 cttttggagt acgtcgtctt taggttgggg ggaggggttt tatgcgatgg agtttcccca
     1021 cactgagtgg gtggagactg aagttaggcc agcttggcac ttgatgtaat tctccttgga
     1081 atttgccctt tttgagtttg gatcttggtt cattctcaag cctcagacag tggttcaaag
     1141 tttttttctt ccatttcagg tgtcgtgaac acgtggccac cgccaccatg aggcggcgct
     1201 cgaggatgct gctctgcttc gccttcctgt gggtgctggg aatcgcctac tacatgtact
     1261 cgggaggagg atctgcgctg gccggaggag cgggaggagg agccggcagg aaggaggact
     1321 ggaatgaaat tgaccccatt aaaaagaaag accttcatca cagcaatgga gaagagaaag
     1381 cacaaagcat ggagaccctc cctccaggga aagtacggtg gccagacttt aaccaggaag
     1441 cttatgttgg agggacgatg gtccgctccg ggcaggaccc ttacgcccgc aacaagttca
     1501 accaggtgga gagtgataag cttcgaatgg atagagccat ccctgacacc cggcatgacc
     1561 agtgtcagcg gaagcagtgg cgggtggatc tgccggccac cagcgtggtg atcacgtttc
     1621 acaatgaagc caggtcggcc ctactcagga ccgtggtcag cgtgcttaag aaaagcccgc
     1681 cccatctcat aaaagaaatc atcttggtgg atgactacag caatgatcct gaggacgggg
     1741 ctctcttggg gaaaattgag aaagtgcgag ttcttagaaa tgatcgacga gaaggcctca
     1801 tgcgctcacg ggttcggggg gccgatgctg cccaagccaa ggtcctgacc ttcctggaca
     1861 gtcactgcga gtgtaatgag cactggctgg agcccctcct ggaaagggtg gcggaggaca
     1921 ggactcgggt tgtgtcaccc atcatcgatg tcattaatat ggacaacttt cagtatgtgg
     1981 gggcatctgc tgacttgaag ggcggttttg attggaactt ggtattcaag tgggattaca
     2041 tgacgcctga gcagagaagg tcccggcagg ggaacccagt cgcccctata aaaaccccca
     2101 tgattgctgg tgggctgttt gtgatggata agttctattt tgaagaactg gggaagtacg
     2161 acatgatgat ggatgtgtgg ggaggagaga acctagagat ctcgttccgc gtgtggcagt
     2221 gtggtggcag cctggagatc atcccgtgca gccgtgtggg acacgtgttc cggaagcagc
     2281 acccctacac gttcccgggt ggcagtggca ctgtctttgc ccgaaacacc cgccgggcag
     2341 cagaggtctg gatggatgaa tacaaaaatt tctattatgc agcagtgcct tctgctagaa
     2401 acgttcctta tggaaatatt cagagcagat tggagcttag gaagaaactc agctgcaagc
     2461 ctttcaaatg gtaccttgaa aatgtctatc cagagttaag ggttccagac catcaggata
     2521 tagcttttgg ggccttgcag cagggaacta actgcctcga cactttggga cactttgctg
     2581 atggtgtggt tggagtttat gaatgtcaca atgctggggg aaaccaggaa tgggccttga
     2641 cgaaggagaa gtcggtgaag cacatggatt tgtgccttac tgtggtggac cgggcaccgg
     2701 gctctcttat aaagctgcag ggctgccgag aaaatgacag cagacagaaa tgggaacaga
     2761 tcgagggcaa ctccaagctg aggcacgtgg gcagcaacct gtgcctggac agtcgcacgg
     2821 ccaagagcgg gggcctaagc gtggaggtgt gtggcccggc cctttcgcag cagtggaagt
     2881 tcacgctcaa cctgcagcag ggaggaagcg gaggaagcat ggatagcact gagagcctgt
     2941 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     3001 gcgtgcgcgg cgagggcgag ggcgatgcca ccaacggcaa gctgaccctg aagttcatct
     3061 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg
     3121 tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     3181 tgcccgaagg ctacgtccag gagcgcacca tcaccttcaa ggacgacggc acctacaaga
     3241 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     3301 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
     3361 acaacgtcta tatcaccgcc gacaagcaga agaacggcat caaggccaac ttcaagatcc
     3421 gccacaacgt ggaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca
     3481 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag tccaagctga
     3541 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     3601 ggatcactgg aggcagtggc ggttcttagt ctagaagctc gctgatcagc ctcgactgtg
     3661 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     3721 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     3781 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     3841 gacaatagca ggcatgctgg ggatggcccg ggctctatgg cttctgaggc ggaaagaacc
     3901 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     3961 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     4021 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     4081 gggctccctt tagggttccg atttagtgct ttacggcacc tctcccccaa aaaacttgat
     4141 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     4201 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     4261 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     4321 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     4381 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4441 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4501 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     4561 ctccgcccag ttccgcccat tctccgcccc taggctgact aatttttttt atttatgcag
     4621 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     4681 gcctaggctt ttgcaaaaag ctcccccccc gggaggtcca caatggttga acaagatgga
     4741 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     4801 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     4861 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tccaggacga ggcagcgcgg
     4921 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     4981 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     5041 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     5101 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     5161 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     5221 ccagccgaac tgttcgccag gctcaaggcg cgtatgcccg acggcgagga tctcgtcgtg
     5281 actcatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     5341 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     5401 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     5461 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     5521 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     5581 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     5641 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     5701 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     5761 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     5821 gtataccgtc gatctttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     5881 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     5941 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     6001 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     6061 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     6121 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     6181 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     6241 gtaggtcgtt cgctccaagc tgggctgtat gcacgaaccc cccgttcagc ccgaccgctg
     6301 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     6361 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     6421 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     6481 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     6541 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     6601 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     6661 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     6721 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     6781 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     6841 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     6901 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     6961 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     7021 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     7081 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     7141 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     7201 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     7261 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     7321 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     7381 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     7441 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     7501 gatgtaaccc actcgggcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     7561 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     7621 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     7681 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     7741 cacatttccc cgaaaagtgc cacctgacgt cagatcgacg gatcgggaga tcggtaccga
     7801 attcaagctt
//
