LOCUS       sequence-333114-        6991 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the nucleoporin Nup62 with C-terminal Flag and EPEA tags. 
            Proteins of interest can be inserted by replacing Nup62 using Sgfl +
            Sgsl restriction enzymes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6991)
  AUTHORS   Ramirez DH, Aonbangkhen C, Wu HY, Naftaly JA, Tang S, O'Meara TR, 
            Woo CM
  TITLE     Engineering a Proximity-Directed O-GlcNAc Transferase for Selective 
            Protein O-GlcNAcylation in Cells.
  JOURNAL   ACS Chem Biol. 2020 Apr 17;15(4):1059-1066. doi: 
            10.1021/acschembio.0c00074. Epub 2020 Mar 2.
  PUBMED    32119511
REFERENCE   2  (bases 1 to 6991)
  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"; doi: 
            "10.1021/acschembio.0c00074"; journalName: "ACS Chem Biol"; date: 
            "2020-04-17- 17"; volume: "15"; issue: "4"; pages: "1059-1066"
COMMENT     Sequence Label: (133) pcDNA3.1-Nup62-Flag-EPEA
FEATURES             Location/Qualifiers
     source          1..6991
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(84..104)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        89..418
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      269..404
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     331..350
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             485..1279
                     /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(539..558)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1149..1168
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1453..1574
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1490..1509)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1544..1563
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1623..1639)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1623..1639)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1636..1658)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1647..1663
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1671..1677,1678..1695,1696..1701))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1716..1737
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1854..1871)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2025..2613)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2105..2124)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2784..3575,3576..3644))
                     /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     3407..3426
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3645..3749)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3824..3843)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4015..4394
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4395..4598
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4549..4569
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4643..4662
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4643..4661
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             6286..6309
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(6362..6379)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6368..6592
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(6638..6991,1..75)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6725..6744)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6935..6956
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
       61 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
      121 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
      181 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
      241 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
      301 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
      361 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
      421 gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt
      481 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
      541 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
      601 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
      661 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
      721 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
      781 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
      841 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
      901 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
      961 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     1021 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     1081 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     1141 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     1201 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     1261 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     1321 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     1381 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     1441 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     1501 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     1561 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     1621 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     1681 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     1741 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     1801 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     1861 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     1921 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     1981 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     2041 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     2101 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     2161 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     2221 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     2281 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     2341 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     2401 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     2461 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     2521 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     2581 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     2641 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     2701 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     2761 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     2821 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     2881 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     2941 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     3001 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     3061 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     3121 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     3181 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     3241 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     3301 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     3361 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     3421 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     3481 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     3541 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     3601 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     3661 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     3721 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     3781 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
     3841 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
     3901 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
     3961 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
     4021 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
     4081 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     4141 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     4201 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     4261 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     4321 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     4381 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
     4441 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
     4501 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
     4561 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
     4621 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
     4681 gtttaaactt aagcttgggc gatcgcaatg gcaagcgggt ttaattttgg aggcactggg
     4741 gcccctacag gcgggttcac gtttggcact gcaaagacgg caacaaccac acctgctaca
     4801 gggttttctt tctccacctc tggcactgga gggtttaatt ttggggctcc cttccaacca
     4861 gccacaagta ccccttccac cggcctgttc tcacttgcca cccagactcc ggccacacag
     4921 acgacaggct tcacttttgg aacagcgact cttgcttcgg ggggaactgg attttctttg
     4981 gggatcggtg cttcaaagct caacttgagc aacacagctg ccaccccagc catggcaaac
     5041 cccagcggct ttgggctggg cagcagcaac ctcactaatg ccatatcgag caccgtcacc
     5101 tccagccagg gcacagcacc caccggcttt gtgtttggcc cctccaccac ctctgtggct
     5161 ccagctacca cacctggagg cttctcattc actggtggaa gcacggccca accctccggt
     5221 ttcaacattg gctcagcagg gaattcagcc cagcccacgg cacctgccac gttgcccttc
     5281 actccggcca cgccagcagc caccacagca ggtgccacac agccagctgc tcccacaccc
     5341 acagccacca tcaccagtac tgggcccagc ctctttgcgt caatagcaac tgctccaacc
     5401 tcatctgcca ccactggact ctccctctgt acccctgtga ccacagcggg cgcccccact
     5461 gctgggacac agggcttcag cttaaaggca cctggagcag cttccggcac ctccacaaca
     5521 acatccaccg ctgccaccgc caccgccacc accaccacca gcagcagcac caccggcttt
     5581 gccttgaatt taaaaccact ggcgccagcc gggatcccca gcaatacagc agctgccgtg
     5641 accgctccac ctggccctgg cgcagctgca ggggcggctg ccagctccgc catgacctac
     5701 gcgcagctgg agagcctgat caacaaatgg agcctggagc tagaggacca ggagcggcac
     5761 ttcctccagc aggccaccca ggtcaacgcc tgggaccgca cgctgatcga gaatggagaa
     5821 aagatcacca gcctgcaccg cgaggtggag aaggtgaagc tggaccagaa gaggctggac
     5881 caggagctcg acttcatcct gtcccagcag aaggagctgg aagacctgct gagcccactg
     5941 gaggagttgg tcaaggagca gagcgggacc atctacctgc agcacgcgga tgaggagcgt
     6001 gagaaaacct acaagctggc tgagaacatc gacgcacagc tcaagcgcat ggcccaggat
     6061 ctcaaggaca tcatcgagca cctgaacacg tccggggccc ccgccgacac cagtgaccca
     6121 ctgcagcaga tctgcaagat cctcaatgcg cacatggact cactgcagtg gatcgaccag
     6181 aactcggccc tgctgcagag gaaggtggag gaggtgacca aggtgtgcga gggccggcgc
     6241 aaggagcagg agcgcagctt ccggatcacc tttgacggcg cgccagacta caaggatgac
     6301 gacgataagg aacctgaagc ataactcgag tctagagggc ccgtttaaac ccgctgatca
     6361 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     6421 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     6481 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     6541 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     6601 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     6661 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     6721 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     6781 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     6841 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     6901 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     6961 acactcaacc ctatctcggt ctattctttt g
//
