LOCUS       sequence-311566-        9547 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Golgi marker ManII-GFP in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9547)
  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 9547)
  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-ManII-GFP
FEATURES             Location/Qualifiers
     source          1..9547
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      96..231
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     158..177
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             271..1065
                     /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(325..344)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     935..954
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1239..1360
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1276..1295)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1330..1349
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1393..1410)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1564..2152)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1644..1663)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2323..3114,3115..3183))
                     /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     2946..2965
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3184..3288)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3364..4526
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          3595..4517
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     4474..4494
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             8014..8721
                     /codon_start=1
                     /product="photostable monomeric superfolder GFP (Valbuena 
                     et al., 2020)"
                     /label=msGFP2
                     /translation="MDSTESLFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTITFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITGGSGGS"
     primer_bind     complement(8056..8077)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     8664..8685
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(8745..8762)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8751..8975
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9021..9449
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9108..9127)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(9458..9478)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(9463..9547,1..245)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
ORIGIN
        1 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
       61 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
      121 ccgcccattc tccgccccta ggctgactaa ttttttttat ttatgcagag gccgaggccg
      181 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
      241 gcaaaaagct ccccccccgg gaggtccaca atggttgaac aagatggatt gcacgcaggt
      301 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
      361 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
      421 accgacctgt ccggtgccct gaatgaactc caggacgagg cagcgcggct atcgtggctg
      481 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
      541 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
      601 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
      661 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
      721 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
      781 ttcgccaggc tcaaggcgcg tatgcccgac ggcgaggatc tcgtcgtgac tcatggcgat
      841 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
      901 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
      961 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     1021 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     1081 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     1141 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     1201 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     1261 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     1321 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     1381 tctttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     1441 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     1501 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     1561 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     1621 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     1681 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     1741 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     1801 ctccaagctg ggctgtatgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     1861 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     1921 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     1981 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     2041 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     2101 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     2161 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     2221 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     2281 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     2341 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     2401 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     2461 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     2521 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     2581 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     2641 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     2701 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     2761 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     2821 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     2881 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     2941 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     3001 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     3061 tcgggcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     3121 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     3181 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     3241 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     3301 aaaagtgcca cctgacgtca gatcgacgga tcgggagatc ggtaccgaat tcaagcttcg
     3361 tgaggctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg
     3421 gggggagggg tcggcaattg aaccggtgcc tagagaaggt ggcgcggggt aaactgggaa
     3481 agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt
     3541 gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acaggtaagt
     3601 gccgtgtgtg gttcccgcgg gcctggcctc tttacgggtt atggcccttg cgtgccttga
     3661 attacttcca cctggctcca gtacgtgatt cttgatcccg agctggagcc aggggcgggc
     3721 cttgcgcttt aggagcccct tcgcctcgtg cttgagttga ggcctggcct gggcgctggg
     3781 gccgccgcgt gcgaatctgg tggcaccttc gcgcctgtct cgctgctttc gataagtctc
     3841 tagccattta aaatttttga tgacctgctg cgacgctttt tttctggcaa gatagtcttg
     3901 taaatgcggg ccaggatctg cacactggta tttcggtttt tggggccgcg ggcggcgacg
     3961 gggcccgtgc gtcccagcgc acatgttcgg cgaggcgggg cctgcgagcg cggccaccga
     4021 gaatcggacg ggggtagtct caagctggcc ggcctgctct ggtgcctggc ctcgcgccgc
     4081 cgtgtatcgc cccgccctgg gcggcaaggc tggcccggtc ggcaccagtt gcgtgagcgg
     4141 aaagatggcc gcttcccggc cctgctccag ggggctcaaa atggaggacg cggcgctcgg
     4201 gagagcgggc gggtgagtca cccacacaaa ggaaaggggc ctttccgtcc tcagccgtcg
     4261 cttcatgtga ctccacggag taccgggcgc cgtccaggca cctcgattag ttctggagct
     4321 tttggagtac gtcgtcttta ggttgggggg aggggtttta tgcgatggag tttccccaca
     4381 ctgagtgggt ggagactgaa gttaggccag cttggcactt gatgtaattc tccttggaat
     4441 ttgccctttt tgagtttgga tcttggttca ttctcaagcc tcagacagtg gttcaaagtt
     4501 tttttcttcc atttcaggtg tcgtgaacac gtggccaccg ccaccatgaa gttaagtcgc
     4561 cagttcaccg tgtttggcag cgcgatcttc tgcgtcgtaa tcttctcact ctacctgatg
     4621 ctggacaggg gtcacttgga ctaccctcgg ggcccgcgcc aggagggctc ctttccgcag
     4681 ggccagcttt caatattgca agaaaagatt gaccatttgg agcgtttgct cgctgagaac
     4741 aacgagatca tctcaaatat cagagactca gtcatcaacc tgagcgagtc tgtggaggac
     4801 ggcccgcggg ggtcaccagg caacgccagc caaggctcca tccacctcca ctcgccacag
     4861 ttggccctgc aggctgaccc cagagactgt ttgtttgctt cacagagtgg gagtcagccc
     4921 cgggatgtgc agatgttgga tgtttacgat ctgattcctt ttgataatcc agatggtgga
     4981 gtttggaagc aaggatttga cattaagtat gaagcggatg agtgggacca tgagcccctg
     5041 caagtgtttg tggtgcctca ctcccataat gacccaggtt ggttgaagac tttcaatgac
     5101 tactttagag acaagactca gtatattttt aataacatgg tcctaaagct gaaagaagac
     5161 tcaagcagga agtttatgtg gtctgagatc tcttaccttg caaaatggtg ggatattata
     5221 gatattccga agaaggaagc tgttaaaagt ttactacaga atggtcagct ggaaattgtg
     5281 accggtggct gggttatgcc tgatgaagcc actccacatt attttgcctt aattgaccaa
     5341 ctaattgaag ggcaccaatg gctggaaaaa aatctaggag tgaaacctcg atcgggctgg
     5401 gccatagatc cctttggtca ttcacccaca atggcttatc ttctaaagcg tgctggattt
     5461 tcacacatgc tcatccagag agtccattat gcaatcaaaa aacacttctc tttgcataaa
     5521 acgctggagt ttttctggag acagaattgg gatcttggat ctgctacaga cattttgtgc
     5581 catatgatgc ccttctacag ctacgacatc cctcacacct gtgggcctga tcctaaaata
     5641 tgctgccagt ttgattttaa acggcttcct ggaggcagat atggttgtcc ctggggagtt
     5701 cccccagaag caatatctcc tggaaatgtc caaagcaggg ctcagatgct attggatcag
     5761 taccggaaaa agtcaaaact tttccgcact aaagttctgc tggctccact gggagacgac
     5821 tttcggttca gtgaatacac agagtgggat ctgcagtgca ggaactacga gcaactgttc
     5881 agttacatga actcgcagcc tcatctgaaa gtgaagatcc agtttggaac cttgtcagat
     5941 tatttcgacg cattggagaa agcggtggca gccgagaaga agagtagcca gtctgtgttc
     6001 cctgccctga gtggagactt cttcacgtac gctgacagag acgaccatta ctggagtggc
     6061 tacttcacgt ccagaccttt ctacaaacga atggacagaa taatggaatc tcgtataagg
     6121 gctgctgaaa ttctttacca gttggccttg aaacaagctc agaaatacaa gataaataaa
     6181 tttctttcat cacctcatta cacaacactg acagaagcca gaaggaactt aggactattt
     6241 cagcatcatg atgccatcac aggaaccgcg aaagactggg tggttgtgga ctatggtacc
     6301 agactctttc agtcattaaa ttctttggag aagataattg gagattctgc atttcttctc
     6361 attttaaagg acaaaaagct gtaccagtca gatccttcca aagccttctt agagatggat
     6421 acgaagcaaa gttcacaaga ttctctgccc caaaaaatta taatacaact gagcgcacag
     6481 gagccaaggt accttgtggt ctacaatccc tttgaacaag aacggcattc agtggtgtcc
     6541 atccgggtaa actccgccac agtgaaagtg ctgtctgatt cgggaaaacc ggtggaggtt
     6601 caagtcagtg cagtttggaa cgacatgagg acaatttcac aagcagccta tgaggtttct
     6661 tttctagctc atataccacc actgggactg aaagtgttta agatcttaga gtcacaaagt
     6721 tcaagctcac acttggctga ttatgtccta tataataatg atggactagc agaaaatgga
     6781 atattccacg tgaagaacat ggtggatgct ggagatgcca taacaataga gaatcccttc
     6841 ctggcgattt ggtttgaccg atctgggctg atggagaaag tgagaaggaa agaagacagt
     6901 agacagcatg aactgaaggt ccagttcctg tggtacggaa ccaccaacaa aagggacaag
     6961 agcggtgcct acctcttcct gcctgacggg cagggccagc catatgtttc cctaagaccg
     7021 ccctttgtca gagtgacacg tggaaggatc tactcagatg tgacctgttt cctcgaacac
     7081 gttactcaca aagtccgcct gtacaacatt cagggaatag aaggtcagtc catggaagtt
     7141 tctaatattg taaacatcag gaatgtgcat aaccgtgaga ttgtaatgag aatttcatct
     7201 aaaataaaca accaaaatag atattatact gacctaaatg gatatcagat tcagcctaga
     7261 aggaccatga gcaaattgcc tcttcaagcc aacgtttacc cgatgtgcac aatggcgtat
     7321 atccaggatg ctgagcaccg gctcacgctg ctctctgctc agtctctagg tgcttccagc
     7381 atggcttctg gtcagattga agtcttcatg gatcgaaggc tcatgcagga tgataaccgt
     7441 ggccttgggc aaggcgtcca tgacaataag attacagcta atttgtttcg aatcctcctc
     7501 gagaagagaa gcgctgtgaa catggaagaa gaaaagaaga gccctgtcag ctacccttcc
     7561 ctcctcagcc acatgacttc gtccttcctc aaccatccct ttctccccat ggtactaagt
     7621 ggccagctcc cctcccctgc ctttgagctg ctgagtgaat ttcctctgct gcagtcctct
     7681 ctaccttgtg atatccatct ggtcaacctg cggacaatac aatcaaagat gggcaaaggc
     7741 tattcggatg aggcagcctt gatcctccac aggaaagggt ttgattgcca gttctccagc
     7801 agaggcatcg ggctaccctg ttccactact cagggaaaga tgtcagttct gaaacttttc
     7861 aacaagtttg ctgtggagag tctcgtccct tcctctctgt ccttgatgca ctcccctcca
     7921 gatgcccaga acatgagtga agtcagcctg agccccatgg agatcagcac gttccgtatc
     7981 cgcttgcgtt ggaccggagg aagcggagga agcatggata gcactgagag cctgttcacc
     8041 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     8101 cgcggcgagg gcgagggcga tgccaccaac ggcaagctga ccctgaagtt catctgcacc
     8161 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag
     8221 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     8281 gaaggctacg tccaggagcg caccatcacc ttcaaggacg acggcaccta caagacccgc
     8341 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     8401 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     8461 gtctatatca ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa gatccgccac
     8521 aacgtggagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     8581 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccaa gctgagcaaa
     8641 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     8701 actggaggca gtggcggttc ttagtctaga agctcgctga tcagcctcga ctgtgccttc
     8761 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     8821 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     8881 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     8941 tagcaggcat gctggggatg gcccgggctc tatggcttct gaggcggaaa gaaccagctg
     9001 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     9061 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     9121 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     9181 ccctttaggg ttccgattta gtgctttacg gcacctctcc cccaaaaaac ttgattaggg
     9241 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     9301 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     9361 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     9421 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     9481 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     9541 gcaacca
//
