LOCUS       sequence-322672-        8283 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GFP tagged Man1B1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8283)
  AUTHORS   Sun X, Mahajan D, Chen B, Song Z, Lu L
  TITLE     A quantitative study of the Golgi retention of glycosyltransferases.
  JOURNAL   J Cell Sci. 2021 Oct 15;134(20). pii: 272560. doi: 
            10.1242/jcs.258564. Epub 2021 Oct 21.
  PUBMED    34533190
REFERENCE   2  (bases 1 to 8283)
  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: "J Cell 
            Sci."; date: "2021-10-15"; volume: "134(20). pii: 272560. doi"; 
            pages: " 10.1242/jcs.258564. Epub 2021 Oct 21"
COMMENT     Sequence Label: Man1B1-GFP
FEATURES             Location/Qualifiers
     source          1..8283
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..729
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     680..700
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     726..750
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          890..1022
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1005..1024
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1067..1085
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1097..1106
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3224..3226,3227..3940)
                     /codon_start=1
                     /product="monomeric GFP variant optimized for use in 
                     oxidizing environments such as the secretory pathway 
                     (Costantini et al., 2015)"
                     /label=moxGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FISTTGKLPVPWPTLVTTLTYGVQSFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(3266..3287)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     polyA_signal    3987..4108
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3998..4017)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4052..4071
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      4294..4749
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4381..4400)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4591..4612
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4866..5223
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4866..5057
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4889..4909)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5074..5209
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5274..6068
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5328..5347)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5938..5957
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6132..6180
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(6221..6240)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6340..6362
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6400..6418)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6486..6590
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6591..6659,6660..7451)
                     /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     complement(6809..6828)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7622..8210
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8111..8130
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactg
      661 cgatcgcccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      721 agcagagctc gtttagtgaa ccgtcagatc actagaagct ttattgcggt agtttatcac
      781 agttaaattg ctaacgcagt cagtgcttct gacacaacag tctcgaactt aagctgcagt
      841 gactctctta aggtagcctt gcagaagttg gtcgtgaggc actgggcagg taagtatcaa
      901 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact
      961 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac
     1021 aggtgtccac tcccagttca attacagctc ttaaggctag agtacttaat acgactcact
     1081 ataggctagc ctcgaggcca ccatggctgc ctgcgagggc aggagaagcg gagctctcgg
     1141 ttcctctcag tcggacttcc tgacgccgcc agtgggcggg gccccttggg ccgtcgccac
     1201 cactgtagtc atgtacccac cgccgccgcc gccgcctcat cgggacttca tctcggtgac
     1261 gctgagcttt ggcgagaact atgacaacag caagagttgg cggcggcgct cgtgctggag
     1321 gaaatggaag caactgtcga gattgcagcg gaatatgatt ctcttcctcc ttgcctttct
     1381 gcttttctgt ggactcctct tctacatcaa cttggctgac cattggaaag ctctggcttt
     1441 caggctagag gaagagcaga agatgaggcc agaaattgct gggttaaaac cagcaaatcc
     1501 acccgtctta ccagctcctc agaaggcgga caccgaccct gagaacttac ctgagatttc
     1561 gtcacagaag acacaaagac acatccagcg gggaccacct cacctgcaga ttagaccccc
     1621 aagccaagac ctgaaggatg ggacccagga ggaggccaca aaaaggcaag aagcccctgt
     1681 ggatccccgc ccggaaggag atccgcagag gacagtcatc agctggaggg gagcggtgat
     1741 cgagcctgag cagggcaccg agctcccttc aagaagagca gaagtgccca ccaagcctcc
     1801 cctgccaccg gccaggacac agggcacacc agtgcatctg aactatcgcc agaagggcgt
     1861 gattgacgtc ttcctgcatg catggaaagg ataccgcaag tttgcatggg gccatgacga
     1921 gctgaagcct gtgtccaggt ccttcagtga gtggtttggc ctcggtctca cactgatcga
     1981 cgcgctggac accatgtgga tcttgggtct gaggaaagaa tttgaggaag ccaggaagtg
     2041 ggtgtcgaag aagttacact ttgaaaagga cgtggacgtc aacctgtttg agagcacgat
     2101 ccgcatcctg ggggggctcc tgagtgccta ccacctgtct ggggacagcc tcttcctgag
     2161 gaaagctgag gattttggaa atcggctaat gcctgccttc agaacaccat ccaagattcc
     2221 ttactcggat gtgaacatcg gtactggagt tgcccacccg ccacggtgga cctccgacag
     2281 cactgtggcc gaggtgacca gcattcagct ggagttccgg gagctctccc gtctcacagg
     2341 ggataagaag tttcaggagg cagtggagaa ggtgacacag cacatccacg gcctgtctgg
     2401 gaagaaggat gggctggtgc ccatgttcat caatacccac agtggcctct tcacccacct
     2461 gggcgtattc acgctgggcg ccagggccga cagctactat gagtacctgc tgaagcagtg
     2521 gatccagggc gggaagcagg agacacagct gctggaagac tacgtggaag ccatcgaggg
     2581 tgtcagaacg cacctgctgc ggcactccga gcccagtaag ctcacctttg tgggggagct
     2641 tgcccacggc cgcttcagtg ccaagatgga ccacctggtg tgcttcctgc cagggacgct
     2701 ggctctgggc gtctaccacg gcctgcccgc cagccacatg gagctggccc aggagctcat
     2761 ggagacttgt taccagatga accggcagat ggagacgggg ctgagtcccg agatcgtgca
     2821 cttcaacctt tacccccagc cgggccgtcg ggacgtggag gtcaagccag cagacaggca
     2881 caacctgctg cggccagaga ccgtggagag cctgttctac ctgtaccgcg tcacagggga
     2941 ccgcaaatac caggactggg gctgggagat tctgcagagc ttcagccgat tcacacgggt
     3001 cccctcgggt ggctattctt ccatcaacaa tgtccaggat cctcagaagc ccgagcctag
     3061 ggacaagatg gagagcttct tcctggggga gacgctcaag tatctgttct tgctcttctc
     3121 cgatgaccca aacctgctca gcctggacgc ctacgtgttc aacaccgaag cccaccctct
     3181 gcctatctgg acccctgcct ctagagtcga ccccgggccg cgggtgtcca agggcgagga
     3241 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     3301 gttctccgtg cggggcgagg gcgagggcga tgccaccaac ggcaagctga ccctgaagtt
     3361 catcagcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
     3421 cggcgtgcag agcttctccc gctaccccga ccacatgaag cgccacgact tcttcaagag
     3481 cgccatgccc gaaggctacg tccaggagcg caccatctcc ttcaaggacg acggcaccta
     3541 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     3601 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaacttcaa
     3661 ctcccacaac gtctatatca ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa
     3721 gatccgccac aacgtggagg acggctccgt gcagctcgcc gaccactacc agcagaacac
     3781 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgtcca cccagtccaa
     3841 gctgtccaaa gaccccaacg agaagcgcga tcacatggtc cttctggaat tcgtgaccgc
     3901 cgccgggatc actcacggca tggacgagct gtacaagtaa gcggccgctt ccctttagtg
     3961 agggttaatg cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac
     4021 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     4081 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     4141 ggttcagggg gagatgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtaa
     4201 aatccgataa ggatcgatcc gggctggcgt aatagcgaag aggcccgcac cgatcgccct
     4261 tcccaacagt tgcgcagcct gaatggcgaa tggacgcgcc ctgtagcggc gcattaagcg
     4321 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     4381 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     4441 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     4501 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     4561 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     4621 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     4681 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
     4741 ttacaatttc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     4801 acgcggatct gcgcagcacc atggcctgaa ataacctctg aaagaggaac ttggttaggt
     4861 accttctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     4921 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     4981 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     5041 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     5101 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     5161 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     5221 aaaaagcttg attcttctga cacaacagtc tcgaacttaa ggctagagcc accatgattg
     5281 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     5341 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     5401 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     5461 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     5521 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     5581 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     5641 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     5701 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     5761 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     5821 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     5881 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     5941 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     6001 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     6061 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     6121 acgatggccg caataaaata tctttatttt cattacatct gtgtgttggt tttttgtgtg
     6181 aatcgatagc gataaggatc cgcgtatggt gcactctcag tacaatctgc tctgatgccg
     6241 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     6301 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     6361 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     6421 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     6481 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     6541 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     6601 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     6661 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     6721 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     6781 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     6841 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     6901 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     6961 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     7021 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     7081 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     7141 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     7201 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     7261 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     7321 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     7381 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     7441 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     7501 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     7561 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     7621 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     7681 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     7741 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     7801 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     7861 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     7921 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     7981 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     8041 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     8101 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     8161 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     8221 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg gctcgacaga
     8281 tct
//
