LOCUS       sequence-358819-        6926 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For transient expression of C-terminal mRuby2-tagged MACROD1del1-77.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6926)
  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 6926)
  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: pcDNA3-mRuby2-MACROD1_d77
FEATURES             Location/Qualifiers
     source          1..6926
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      911..920
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1642..1671
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="ASMTGGQQMG"
     CDS             join(1675..1683,1684..1698)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             1705..2415
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH
                     MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV
                     AKFAGLGGGMDELYK"
     promoter        complement(2479..2497)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2480..2497)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2517..2534)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2523..2747
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2793..3221
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2880..2899)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3090..3111
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3230..3250)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3235..3564
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3415..3550
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3477..3496
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3631..4425
                     /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(3685..3704)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4295..4314
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4599..4720
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4636..4655)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4690..4709
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4769..4785)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4769..4785)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4782..4804)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4793..4809
                     /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(4817..4823,4824..4841,4842..4847))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4862..4883
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5000..5017)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5171..5759)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5251..5270)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5930..6721,6722..6790))
                     /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     6553..6572
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6791..6895)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttatgtgg
      901 gcccccctgg ccatggcggc gaaggtggac ctgagcacct ccaccgactg gaaggaggcg
      961 aaatcctttc tgaagggcct gagtgacaag cagcgggagg aacattactt ctgcaaggac
     1021 tttgtcaggc tgaagaagat cccgacatgg aaggagatgg cgaaaggggt ggctgtgaag
     1081 gtggaggagc ccaggtataa aaaggacaag cagctcaatg agaaaatctc cctgctccgc
     1141 agcgacatca ccaagctgga ggtggacgcc atcgtcaacg ccgccaacag ctccctgctc
     1201 ggaggcggtg gcgtggacgg ctgcattcat cgggccgccg gccccctgct taccgacgag
     1261 tgccggaccc tgcagagctg taagactggc aaggccaaga tcaccggcgg ctatcggctc
     1321 ccggccaagt acgtcatcca cacagtgggg cccatcgcct acggggagcc cagcgccagc
     1381 caggctgccg agctccgcag ctgctacctg agcagtctgg acctgctgct ggagcaccgg
     1441 ctccgctcgg tggcgttccc ctgcatctcc accggcgtgt ttggctaccc ctgtgaggcg
     1501 gccgccgaga tcgtgctggc cacgctgcga gagtggctgg agcagcacaa ggacaaggtg
     1561 gaccggctga tcatctgcgt gttcctcgag aaggacgagg acatctaccg gagccggctc
     1621 ccccactact tccccgtggc cgctagcatg actggtggac agcaaatggg tcgggatctg
     1681 tacgacgatg acgataagga tccgatggtg tctaagggcg aagagctgat caaggaaaat
     1741 atgcgtatga aggtggtcat ggaaggttcg gtcaacggcc accaattcaa atgcacaggt
     1801 gaaggagaag gcaatccgta catgggaact caaaccatga ggatcaaagt catcgaggga
     1861 ggacccctgc catttgcctt tgacattctt gccacgtcgt tcatgtatgg cagccgtact
     1921 tttatcaagt acccgaaagg cattcctgat ttctttaaac agtcctttcc tgagggtttt
     1981 acttgggaaa gagttacgag atacgaagat ggtggagtcg tcaccgtcat gcaggacacc
     2041 agccttgagg atggctgtct cgtttaccac gtccaagtca gaggggtaaa ctttccctcc
     2101 aatggtcccg tgatgcagaa gaagaccaag ggttgggagc ctaatacaga gatgatgtat
     2161 ccagcagatg gtggtctgag gggatacact catatggcac tgaaagttga tggtggtggc
     2221 catctgtctt gctctttcgt aacaacttac aggtcaaaaa agaccgtcgg gaacatcaag
     2281 atgcccggta tccatgccgt tgatcaccgc ctggaaaggt tagaggaaag tgacaatgaa
     2341 atgttcgtag tacaacgcga acacgcagtt gccaagttcg ccgggcttgg tggtgggatg
     2401 gacgagctgt acaagtaaga attctgcaga tatccatcac actggcggcc gctcgagcat
     2461 gcatctagag ggccctattc tatagtgtca cctaaatgct agagctcgct gatcagcctc
     2521 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     2581 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     2641 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     2701 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     2761 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     2821 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     2881 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     2941 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     3001 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     3061 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     3121 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     3181 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     3241 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3301 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3361 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     3421 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     3481 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     3541 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     3601 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     3661 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     3721 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     3781 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     3841 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     3901 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     3961 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     4021 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     4081 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     4141 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     4201 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     4261 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     4321 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     4381 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     4441 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     4501 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     4561 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     4621 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     4681 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     4741 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     4801 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     4861 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     4921 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     4981 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     5041 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     5101 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     5161 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     5221 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     5281 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     5341 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     5401 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     5461 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     5521 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     5581 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     5641 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     5701 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     5761 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     5821 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     5881 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     5941 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     6001 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     6061 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     6121 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     6181 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     6241 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     6301 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     6361 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     6421 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     6481 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     6541 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     6601 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     6661 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     6721 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     6781 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     6841 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     6901 ttccccgaaa agtgccacct gacgtc
//
