LOCUS       sequence-322675-        8952 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses myc tagged Man2a1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8952)
  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 8952)
  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: Man2a1-myc
FEATURES             Location/Qualifiers
     source          1..8952
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        140..519
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        520..731
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     682..702
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     728..752
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          892..1024
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1007..1026
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1069..1087
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4549..4578
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4579..4608
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     polyA_signal    4658..4779
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4669..4688)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4723..4742
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      4965..5420
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5052..5071)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5262..5283
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5537..5894
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5537..5728
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5560..5580)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5745..5880
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5945..6739
                     /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(5999..6018)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6609..6628
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6803..6851
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(6892..6911)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7011..7033
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7071..7089)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7157..7261
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7262..7330,7331..8122)
                     /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(7480..7499)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8293..8881
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8782..8801
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cttcaatatt ggccattagc catattattc attggttata tagcataaat caatattggc
       61 tattggccat tgcatacgtt gtatctatat cataatatgt acatttatat tggctcatgt
      121 ccaatatgac cgccatgttg gcattgatta ttgactagtt attaatagta atcaattacg
      181 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
      241 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
      301 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
      361 gcccacttgg cagtacatca agtgtatcat atgccaagtc cgccccctat tgacgtcaat
      421 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttacggga ctttcctact
      481 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
      541 accaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
      601 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
      661 tgcgatcgcc cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
      721 taagcagagc tcgtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc
      781 acagttaaat tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca
      841 gtgactctct taaggtagcc ttgcagaagt tggtcgtgag gcactgggca ggtaagtatc
      901 aaggttacaa gacaggttta aggagaccaa tagaaactgg gcttgtcgag acagagaaga
      961 ctcttgcgtt tctgataggc acctattggt cttactgaca tccactttgc ctttctctcc
     1021 acaggtgtcc actcccagtt caattacagc tcttaaggct agagtactta atacgactca
     1081 ctataggcta gcctcgaggc caccatgaag ttaagccgcc agttcaccgt gttcggcagt
     1141 gcgatcttct gtgtggtgat tttctcgctc tacctgatgc tggaccgggg tcacttagac
     1201 taccccagga acccgcgccg cgagggctcc ttccctcagg gccagctctc aatgttgcaa
     1261 gaaaaaatag accatttgga gcgtttgcta gctgagaata atgagatcat ctcaaatatt
     1321 agagactcag tcatcaattt gagtgagtct gtggaggatg gtccgaaaag ttcacaaagc
     1381 aatttcagcc aaggtgctgg ctcacatctt ctgccctcac aattatccct ctcagttgac
     1441 actgcagact gtctgtttgc ttcacaaagt ggaagtcaca attcagatgt gcagatgttg
     1501 gatgtttaca gtctaatttc ttttgacaat ccagatggtg gagtttggaa gcaaggattt
     1561 gacattactt atgaatctaa tgaatgggac actgaacccc ttcaagtctt tgtggtgcct
     1621 cattcccata acgacccagg ttggttgaag actttcaatg actactttag agacaagact
     1681 cagtatattt ttaataacat ggtcctaaag ctgaaagaag actcacggag gaagtttatt
     1741 tggtctgaga tctcttacct ttcaaagtgg tgggatatta tagatattca gaagaaggat
     1801 gctgttaaaa gtttaataga aaatggtcag cttgaaattg tgacaggtgg ctgggttatg
     1861 cctgatgaag ctactccaca ttattttgcc ttaattgatc aactaattga aggacatcag
     1921 tggctggaaa ataatatagg agtgaaacct cggtccggct gggctattga tccctttgga
     1981 cactcaccaa caatggctta tcttctaaac cgtgctggac tttctcacat gcttatccag
     2041 agagttcatt atgcagttaa aaaacacttt gcactgcata aaacattgga gtttttttgg
     2101 agacagaatt gggatctggg atctgtcaca gatattttat gccacatgat gcccttctac
     2161 agctatgaca tccctcacac ttgtggacct gatcctaaaa tatgctgcca gtttgatttt
     2221 aaacgtcttc ctggaggcag atttggttgt ccctggggag tccccccaga aacaatacat
     2281 cctggaaatg tccaaagcag ggctcggatg ctactagatc agtaccgaaa gaagtcaaag
     2341 ctttttcgta ccaaagttct cctggctcca ctaggagatg atttccgcta ctgtgaatac
     2401 acggaatggg atttacagtt taagaattat cagcagcttt ttgattatat gaattctcag
     2461 tccaagttta aagttaagat acagtttgga actttatcag atttttttga tgcgctggat
     2521 aaagcagatg aaactcagag agacaagggc cagtcgatgt tccctgtttt aagtggagat
     2581 tttttcactt atgccgatcg agatgatcat tactggagtg gctattttac atccagaccc
     2641 ttttacaaac gaatggacag aatcatggaa tctcatttaa gggctgctga aattctttac
     2701 tatttcgccc tgagacaagc tcacaaatac aagataaata aatttctctc atcatcactt
     2761 tacacggcac tgacagaagc cagaaggaat ttgggactgt ttcaacatca tgatgctatc
     2821 acaggaactg caaaagactg ggtggttgtg gattatggta ccagaccttt tcattcgtta
     2881 atggttttgg agaagataat tggaaattct gcatttcttc ttattttgaa ggacaaactc
     2941 acatacgact cttactctcc tgataccttc ctggagatgg atttgaaaca aaaatcacaa
     3001 gattctctgc cacaaaaaaa tataataagg ctgagtgcgg agccaaggta ccttgtggtc
     3061 tataatcctc tagaacaaga ccgaatctcg ttggtctcag tctatgtgag ttccccgaca
     3121 gtgcaagtgt tctctgcttc aggaaaacct gtggaagttc aagtcagcgc agtttgggat
     3181 acagcaaata ctatttcaga aacagcctat gagatctctt ttcgagcaca tataccgcca
     3241 ttgggactga aagtgtataa gattttggaa tcagcaagtt caaattcaca tttagctgat
     3301 tatgtcttgt ataagaataa agtagaagat agcggaattt tcaccataaa gaatatgata
     3361 aatactgaag aaggtataac actagagaac tcctttgttt tacttcggtt tgatcaaact
     3421 ggacttatga agcaaatgat gactaaagaa gatggtaaac accatgaagt aaatgtgcaa
     3481 ttttcatggt atggaaccac aattaaaaga gacaaaagtg gtgcctacct cttcttacct
     3541 gatggtaatg ccaagcctta tgtttacaca acaccgccct ttgtcagagt gacacatgga
     3601 aggatttatt cggaagtgac ttgctttttt gaccatgtta ctcatagagt ccgactatac
     3661 cacatacagg gaatagaagg acagtctgtg gaagtttcca atattgtgga catccgaaaa
     3721 gtatataacc gtgagattgc aatgaaaatt tcttctgata taaaaagcca aaatagattt
     3781 tatactgacc taaatgggta ccagattcaa cctagaatga cactgagcaa attgcctctt
     3841 caagcaaatg tctatcccat gaccacaatg gcctatatcc aggatgccaa acatcgtttg
     3901 acactgctct ctgctcagtc tttaggggtt tcgagtttga atagtggtca gattgaagtt
     3961 atcatggatc gaagactcat gcaagatgat aatcgtggcc ttgagcaagg tatccaggat
     4021 aacaagatta cagctaatct atttcgaata ctactagaaa aaagaagtgc tgttaatacg
     4081 gaagaagaaa agaagtcggt cagttatcct tctctcctta gccacataac ttcttctctc
     4141 atgaatcatc cagtcattcc aatggcaaat aagttctcac ctacccttga gctgcaaggt
     4201 gaattctctc cattacagtc atctttgcct tgtgacattc atctggttaa tttgagaaca
     4261 atacagtcaa aggtgggcaa tgggcactcc aatgaggcag ccttgatcct ccacagaaaa
     4321 gggtttgatt gtcggttctc tagcaaaggc acagggctgt tttgttctac tactcaggga
     4381 aagatattgg tacagaaact tttaaacaag tttattgtcg aaagtctcac accttcatca
     4441 ctatccttga tgcattcacc tcccggcact cagaatataa gtgagatcaa cttgagtcca
     4501 atggaaatca gcacattccg aatccagttg aggccgcggg gccggccaga acaaaaactc
     4561 atctcagaag aggatctgga acaaaaactc atctcagaag aggatctgta agcggccgct
     4621 tccctttagt gagggttaat gcttcgagca gacatgataa gatacattga tgagtttgga
     4681 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     4741 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     4801 tttatgtttc aggttcaggg ggagatgtgg gaggtttttt aaagcaagta aaacctctac
     4861 aaatgtggta aaatccgata aggatcgatc cgggctggcg taatagcgaa gaggcccgca
     4921 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggacgcgc cctgtagcgg
     4981 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     5041 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     5101 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     5161 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     5221 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     5281 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     5341 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     5401 aatattaacg cttacaattt cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     5461 tcacaccgca tacgcggatc tgcgcagcac catggcctga aataacctct gaaagaggaa
     5521 cttggttagg taccttctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt
     5581 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     5641 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     5701 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     5761 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     5821 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     5881 taggcttttg caaaaagctt gattcttctg acacaacagt ctcgaactta aggctagagc
     5941 caccatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     6001 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     6061 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     6121 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     6181 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     6241 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     6301 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     6361 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     6421 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     6481 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     6541 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     6601 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     6661 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     6721 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     6781 aacctgccat cacgatggcc gcaataaaat atctttattt tcattacatc tgtgtgttgg
     6841 ttttttgtgt gaatcgatag cgataaggat ccgcgtatgg tgcactctca gtacaatctg
     6901 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     6961 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     7021 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     7081 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     7141 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     7201 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     7261 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     7321 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     7381 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     7441 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     7501 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     7561 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     7621 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     7681 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     7741 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     7801 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     7861 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     7921 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     7981 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     8041 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     8101 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     8161 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     8221 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     8281 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     8341 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     8401 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     8461 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     8521 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     8581 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     8641 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     8701 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     8761 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     8821 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     8881 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     8941 ggctcgacag at
//
