LOCUS       sequence-358811-        6070 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For transient expression of N-terminal GFP-tagged MACROD2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6070)
  AUTHORS   Zaja R, Aydin G, Lippok BE, Feederle R, Luscher B, Feijs KLH
  TITLE     Comparative analysis of MACROD1, MACROD2 and TARG1 expression, 
            localisation and interactome.
  JOURNAL   Sci Rep. 2020 May 19;10(1):8286. doi: 10.1038/s41598-020-64623-y.
  PUBMED    32427867
REFERENCE   2  (bases 1 to 6070)
  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: "Sci Rep."; 
            date: "2020-05-19"; volume: "10(1):8286. doi"; pages: " 
            10.1038/s41598-020-64623-y"
COMMENT     Sequence Label: pDEST-EGFP-MACROD2
FEATURES             Location/Qualifiers
     source          1..6070
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        63..366
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        367..570
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     521..541
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      609..618
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(615..617,618..620,621..1331)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(615..617,618..620,621..1331)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(660..681)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(921..940)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1268..1289
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    1395..1419
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2701..2725)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    2860..2981
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2897..2916)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2951..2970
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2988..3443)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3125..3146)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3337..3356
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3470..3574
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3576..3933
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3576..3767
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3599..3619)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3784..3919
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3968..4762
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3968..4762
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4022..4041)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4632..4651
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4950..4969)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4994..5041
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5370..5958
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5859..5878
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 attagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
       61 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
      121 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
      181 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
      241 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
      301 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
      361 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
      421 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
      481 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
      541 gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat ccgctagcgc
      601 taccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
      661 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
      721 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
      781 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
      841 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
      901 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
      961 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     1021 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg
     1081 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca
     1141 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     1201 tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc
     1261 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     1321 agctgtacaa gtccggactc agatctcgag ctcaagcttc gaattctgca gtcgacggta
     1381 ccgcgggccc atcaacaagt ttgtacaaaa aagcaggctt catgtacccc agcaacaaga
     1441 agaaaaaggt gtggagagag gagaaagaac gtttattgaa gatgacctta gaagagagac
     1501 gcaaagaata cctaagagac tatattcccc tgaacagcat tctgtcatgg aaggaggaga
     1561 tgaagggcaa gggccaaaat gatgaagaaa atactcagga aacatcccag gtgaagaaaa
     1621 gtttgactga aaaagtttct ctctatagag gtgacatcac attgctagag gtagatgcta
     1681 tagtcaatgc cgcaaatgcc agtcttcttg gaggaggagg tgtggatggc tgtattcata
     1741 gagcagccgg cccctgtttg ctagctgaat gtcgtaacct gaatggctgt gatactggac
     1801 atgcaaaaat cacatgtggc tatgaccttc ctgcaaaata tgtcatccat actgtagggc
     1861 caatagccag gggccatatt aatggttccc acaaggaaga ccttgcaaat tgctataaat
     1921 catctctgaa gctcgtgaaa gaaaataaca tccgatcagt tgcatttccc tgcatctcaa
     1981 caggcattta tggctttccc aacgagcctg ctgcagtcat tgccctcaac accattaagg
     2041 aatggcttgc caagaatcac catgaggtgg atcggatcat tttctgtgtc ttcttagaag
     2101 ttgacttcaa aatctacaaa aagaaaatga atgagttttt ctccgtagac gataataatg
     2161 aagaagaaga ggatgttgaa atgaaagaag attcagatga gaacggtcca gaggagaagc
     2221 aaagtgtgga agaaatggaa gagcagagcc aagatgcaga tggtgtcaac actgtcactg
     2281 tgcccggccc tgcttcagaa gaggcagttg aagactgtaa agatgaagat tttgcaaagg
     2341 atgaaaatat tacaaaaggc ggtgaagtga cagatcattc tgtgcgtgac caagatcatc
     2401 ccgatggaca agagaatgat tcaacgaaga atgaaataaa aattgaaaca gaatcgcaga
     2461 gctcatatat ggaaacagaa gaactttcat caaaccaaga agatgccgtg attgtggagc
     2521 aaccagaagt gattccatta acagaggacc aagaagaaaa agaaggtgaa aaagctccag
     2581 gcgaggacac acctaggatg cctgggaaaa gtgaaggctc cagtgaccta gaaaatactc
     2641 caggtcctga tgcaggggca caagatgaag cgaaggaaca aagaaatgga actaaatgag
     2701 acccagcttt cttgtacaaa gtggttgatg ggatccaccg gatctagata actgatcata
     2761 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     2821 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     2881 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     2941 cattctagtt gtggtttgtc caaactcatc aatgtatctt aacgcgtaaa ttgtaagcgt
     3001 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     3061 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     3121 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     3181 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     3241 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     3301 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     3361 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     3421 taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     3481 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     3541 ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg
     3601 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3661 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     3721 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     3781 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     3841 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     3901 ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc
     3961 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     4021 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     4081 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     4141 tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     4201 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     4261 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     4321 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     4381 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     4441 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat
     4501 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     4561 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     4621 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     4681 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     4741 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     4801 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     4861 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     4921 ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat accggaagga
     4981 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt
     5041 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     5101 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     5161 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca
     5221 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     5281 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     5341 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     5401 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     5461 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     5521 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     5581 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     5641 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     5701 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     5761 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     5821 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     5881 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     5941 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     6001 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     6061 accgccatgc
//
