LOCUS       sequence-467876-        7086 bp    DNA     circular SYN 15-APR-2025
DEFINITION  mammalian expression (murine Pgk promoter) of murine TDG fused to 
            BioID2-HA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7086)
  TITLE     The TDG Protein Environment Connects Active DNA Demethylation with 
            Chromatin and RNA Biology
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7086)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pPgk-mTDGa-BioID2-HA
FEATURES             Location/Qualifiers
     source          1..7086
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(46..65)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(204..227)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     554..573
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             1927..2622
                     /codon_start=1
                     /product="promiscuous R40G mutant of a biotin protein 
                     ligase from Aquifex aeolicus (Kim et al., 2016)"
                     /label=BioID2
                     /translation="FKNLIWLKEVDSTQERLKEWNVSYGTALVADRQTKGRGGLGRKWL
                     SQEGGLYFSFLLNPKEFENLLQLPLVLGLSVSEALEEITEIPFSLKWPNDVYFQEKKVS
                     GVLCELSKDKLIVGIGINVNQREIPEEIKDRATTLYEITGKDWDRKEVLLKVLKRISEN
                     LKKFKEKSFKEFKGKIESKMLYLGEEVKLLGEGKITGKLVGLSEKGGALILTEEGIKEI
                     LSGEFSLRRS"
     CDS             2623..2649
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2679..2696)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2685..2909
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2955..3383
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3042..3061)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3252..3273
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3392..3412)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3397..3726
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3577..3712
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3639..3658
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3793..4587
                     /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(3847..3866)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4457..4476
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4761..4882
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4798..4817)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4852..4871
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4931..4947)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4931..4947)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4944..4966)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4955..4971
                     /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(4979..4985,4986..5003,5004..5009))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5024..5045
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5162..5179)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5333..5921)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5413..5432)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6092..6883,6884..6952))
                     /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     6715..6734
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6953..7057)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
       61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
      121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattggg gtaggggagg
      181 cgcttttccc aaggcagtct ggagcatgcg ctttagcagc cccgctgggc acttggcgct
      241 acacaagtgg cctctggcct cgcacacatt ccacatccac cggtaggcgc caaccggctc
      301 cgttctttgg tggccccttc gcgccacctt ctactcctcc cctagtcagg aagttccccc
      361 ccgccccgca gctcgcgtcg tgcaggacgt gacaaatgga agtagcacgt ctcactagtc
      421 tcgtgcagat ggacagcacc gctgagcaat ggaagcgggt aggcctttgg ggcagcggcc
      481 aatagcagct ttgctccttc gctttctggg ctcaggggcg gggcgggcgc ccgaaggtcc
      541 tccggaggcc cggcattctg cacgcttcaa aagcgcacgt ctgccgcgct gttctcctct
      601 tcctcatctc cgggcctttc ggctagcatg gacgcagagg ccgcgcgcag ctattctctg
      661 gagcaagttc aagctttgta ttcatttcca tttcaacaaa tgatggcaga agttcctaac
      721 atggcagtca cgactggaca gcaggtgcca gcagtagctc ctaacatggc aaccgtgaca
      781 gaacagcagg tgcccgcaga cgctcctgtc caggaacctg caccagaagc tccaaagaga
      841 aggaaaagga aacccagagc agcagagccc caggaaccag tggagcccaa aaaacctgct
      901 acgtcgaaga aatccggcaa gtctacaaaa tcaaaggaaa agcaggagaa aatcacagac
      961 gcgtttaaag tgaaaaggaa agtggaccgc ttcaacggcg tctctgaagc tgagcttctg
     1021 accaagactc ttcctgacat tttgaccttc aatctggata ttgtgatcat tggcattaac
     1081 ccgggattaa tggctgctta caaaggacat cactaccctg ggcctggaaa tcacttctgg
     1141 aagtgtctgt tcatgtcggg gctgagtgag gtgcagctga atcacatgga tgaccacacc
     1201 ttacccggca agtacggcat cggattcacc aacatggtgg aacggacgac gccgggcagc
     1261 aaggatctgt ctagtaaaga gttccgggaa ggagggcgca tcctggtgca gaaactgcag
     1321 aaatatcagc cacgaatagc ggtgtttaat ggaaaatgta tttatgaaat tttcagtaaa
     1381 gaagtttttg gagtaaaggt taagaacttg gaatttgggc ttcaacccca caagatccca
     1441 gacacagaaa ctctgtgcta cgtcatgccg tcgtccagcg ccagatgtgc tcagtttccc
     1501 cgggcccagg acaaagttca ttactacatt aagctgaagg acttgagaga ccagctgaaa
     1561 ggcattgaac gcaacaccga cgttcaggaa gtgcagtata catttgacct gcagcttgcg
     1621 caagaggacg caaagaagat ggctgttaag gaagaaaagt atgatccagg ctatgaggca
     1681 gcttacggcg gcgcctatgg ggaaaaccca tgtaatgggg aaccttgtgg cattgcttca
     1741 aatgggctaa cagcccacag tgcggagccg agaggagaag cgacccccgg cgatgttcca
     1801 aatgggcagt ggatggcaca gtcgtttgca gagcagatcc cttcttttaa taattgtggg
     1861 acccgagagc aggaagaaga gagccacgct gaccggtcgt acgtctccgg attcgaattc
     1921 ggatccttca agaacctgat ctggctgaag gaggtggaca gcacccagga gagactgaag
     1981 gagtggaacg tgagctacgg caccgccctg gtggccgaca gacagaccaa gggcagaggc
     2041 ggcctgggca gaaagtggct gagccaggag ggcggcctgt acttcagctt cctgctgaac
     2101 cccaaggagt tcgagaacct gctgcagctg cccctggtgc tgggcctgag cgtgagcgag
     2161 gccctggagg agatcaccga gatccccttc agcctgaagt ggcccaacga cgtgtacttc
     2221 caggagaaga aggtgagcgg cgtgctgtgc gagctgagca aggacaagct gatcgtgggc
     2281 atcggcatca acgtgaacca gagagagatc cccgaggaga tcaaggacag agccaccacc
     2341 ctgtacgaga tcaccggcaa ggactgggac agaaaggagg tgctgctgaa ggtgctgaag
     2401 agaatcagcg agaacctgaa gaagttcaag gagaagagct tcaaggagtt caagggcaag
     2461 atcgagagca agatgctgta cctgggcgag gaggtgaagc tgctgggcga gggcaagatc
     2521 accggcaagc tggtgggcct gagcgagaag ggcggcgccc tgatcctgac cgaggagggc
     2581 atcaaggaga tcctgagcgg cgagttcagc ctgagaagaa gctaccctta cgatgtaccg
     2641 gattacgcat agaagcttaa gtttaaaccg ctgatcagcc tcgactgtgc cttctagttg
     2701 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     2761 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     2821 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     2881 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
     2941 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     3001 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     3061 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     3121 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     3181 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     3241 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     3301 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     3361 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     3421 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3481 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3541 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     3601 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     3661 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     3721 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga
     3781 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     3841 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     3901 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     3961 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     4021 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     4081 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     4141 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     4201 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     4261 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     4321 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     4381 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     4441 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     4501 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     4561 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     4621 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     4681 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     4741 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     4801 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     4861 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     4921 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     4981 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     5041 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     5101 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     5161 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     5221 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     5281 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     5341 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     5401 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     5461 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     5521 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     5581 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     5641 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     5701 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     5761 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     5821 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     5881 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     5941 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     6001 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     6061 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     6121 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     6181 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     6241 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     6301 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     6361 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     6421 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     6481 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     6541 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     6601 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     6661 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     6721 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     6781 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     6841 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     6901 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     6961 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     7021 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     7081 ctgacg
//
