LOCUS Exported 8857 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expression of wild-type catalytic domain only form of TET1. ACCESSION . VERSION . KEYWORDS pEF1a_CD WT TET1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8857) AUTHORS Vincenzetti L, Leoni C, Chirichella M, Kwee I, Monticelli S TITLE The contribution of active and passive mechanisms of 5mC and 5hmC removal in human T lymphocytes is differentiation- and activation-dependent. JOURNAL Eur J Immunol. 2019 Jan 30. doi: 10.1002/eji.201847967. PUBMED 30698829 REFERENCE 2 (bases 1 to 8857) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8857 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(45..64) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" promoter 352..1530 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 583..1521 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 1478..1498 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" primer_bind 1547..1566 /label=T7 /note="T7 promoter, forward primer" promoter 1547..1565 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" regulatory 1598..1607 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1607..1630 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 1643..1669 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 4367..4408 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" CDS 4418..4435 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(4458..4475) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 4464..4688 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 4734..5162 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4821..4840) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5031..5052 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(5171..5191) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 5176..5505 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 5356..5491 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5418..5437 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 5572..6366 /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(5626..5645) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 6236..6255 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 6542..6663 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6579..6598) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 6633..6652 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(6712..6728) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6712..6728) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(6725..6747) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 6736..6752 /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(6760..6790) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6805..6826 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(6943..6960) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(7103..7691) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(7183..7202) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7862..8722) /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 8485..8504 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8723..8827) /gene="bla" /label=AmpR promoter ORIGIN 1 cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat 61 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc 121 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg 181 cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgttgacat 241 tgattattga ctaggctttt gcaaaaagct ttgcaaagat ggataaagtt ttaaacagag 301 aggaatcttt gcagctaatg gaccttctag gtcttgaaag gagtgggaat tggctccggt 361 gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc 421 ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg 481 tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc 541 gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt 601 tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat tacttccacc 661 tggctgcagt acgtgattct tgatcccgag cttcgggttg gaagtgggtg ggagagttcg 721 aggccttgcg cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg gcctgggcgc 781 tggggccgcc gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc tttcgataag 841 tctctagcca tttaaaattt ttgatgacct gctgcgacgc tttttttctg gcaagatagt 901 cttgtaaatg cgggccaaga tctgcacact ggtatttcgg tttttggggc cgcgggcggc 961 gacggggccc gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg agcgcggcca 1021 ccgagaatcg gacgggggta gtctcaagct ggccggcctg ctctggtgcc tggcctcgcg 1081 ccgccgtgta tcgccccgcc ctgggcggca aggctggccc ggtcggcacc agttgcgtga 1141 gcggaaagat ggccgcttcc cggccctgct gcagggagct caaaatggag gacgcggcgc 1201 tcgggagagc gggcgggtga gtcacccaca caaaggaaaa gggcctttcc gtcctcagcc 1261 gtcgcttcat gtgactccac ggagtaccgg gcgccgtcca ggcacctcga ttagttctcg 1321 agcttttgga gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat ggagtttccc 1381 cacactgagt gggtggagac tgaagttagg ccagcttggc acttgatgta attctccttg 1441 gaatttgccc tttttgagtt tggatcttgg ttcattctca agcctcagac agtggttcaa 1501 agtttttttc ttccatttca ggtgtcgtga ggaattagct tggtactaat acgactcact 1561 atagggagac ccaagctggc taggtaagct tggtaccgcc accatggact acaaggacga 1621 cgatgacaag ctcgatggag gataccccta cgacgtgccc gactacgccg gaggatatcc 1681 tgaatcagca gaggaaaagg tgaaggttga accattggat tcactcagct tatttcatct 1741 taaaacggaa tccaacggga aggcattcac tgataaagct tataattctc aggtacagtt 1801 aacggtgaat gccaatcaga aagcccatcc tttgacccag ccctcctctc cacctaacca 1861 gtgtgctaac gtgatggcag gcgatgacca aatacggttt cagcaggttg ttaaggagca 1921 actcatgcat cagagactgc caacattgcc tggtatctct catgaaacac ccttaccgga 1981 gtcagcacta actctcagga atgtaaatgt agtgtgttca ggtggaatta cagtggtttc 2041 taccaaaagt gaagaggaag tctgttcatc cagttttgga acatcagaat tttccacagt 2101 ggacagtgca cagaaaaatt ttaatgatta tgccatgaac ttctttacta accctacaaa 2161 aaacctagtg tctataacta aagattctga actgcccacc tgcagctgtc ttgatcgagt 2221 tatacaaaaa gacaaaggcc catattatac acaccttggg gcaggaccaa gtgttgctgc 2281 tgtcagggaa atcatggaga ataggtatgg tcaaaaagga aacgcaataa ggatagaaat 2341 agtagtgtac accggtaaag aagggaaaag ctctcatggg tgtccaattg ctaagtgggt 2401 tttaagaaga agcagtgatg aagaaaaagt tctttgtttg gtccggcagc gtacaggcca 2461 ccactgtcca actgctgtga tggtggtgct catcatggtg tgggatggca tccctcttcc 2521 aatggccgac cggctataca cagagctcac agagaatcta aagtcataca atgggcaccc 2581 taccgacaga agatgcaccc tcaatgaaaa tcgtacctgt acatgtcaag gaattgatcc 2641 agagacttgt ggagcttcat tctcttttgg ctgttcatgg agtatgtact ttaatggctg 2701 taagtttggt agaagcccaa gccccagaag atttagaatt gatccaagct ctcccttaca 2761 tgaaaaaaac cttgaagata acttacagag tttggctaca cgattagctc caatttataa 2821 gcagtatgct ccagtagctt accaaaatca ggtggaatat gaaaatgttg cccgagaatg 2881 tcggcttggc agcaaggaag gtcgtccctt ctctggggtc actgcttgcc tggacttctg 2941 tgctcatccc cacagggaca ttcacaacat gaataatgga agcactgtgg tttgtacctt 3001 aactcgagaa gataaccgct ctttgggtgt tattcctcaa gatgagcagc tccatgtgct 3061 acctctttat aagctttcag acacagatga gtttggctcc aaggaaggaa tggaagccaa 3121 gatcaaatct ggggccatcg aggtcctggc accccgccgc aaaaaaagaa cgtgtttcac 3181 tcagcctgtt ccccgttctg gaaagaagag ggctgcgatg atgacagagg ttcttgcaca 3241 taagataagg gcagtggaaa agaaacctat tccccgaatc aagcggaaga ataactcaac 3301 aacaacaaac aacagtaagc cttcgtcact gccaacctta gggagtaaca ctgagaccgt 3361 gcaacctgaa gtaaaaagtg aaaccgaacc ccattttatc ttaaaaagtt cagacaacac 3421 taaaacttat tcgctgatgc catccgctcc tcacccagtg aaagaggcat ctccaggctt 3481 ctcctggtcc ccgaagactg cttcagccac accagctcca ctgaagaatg acgcaacagc 3541 ctcatgcggg ttttcagaaa gaagcagcac tccccactgt acgatgcctt cgggaagact 3601 cagtggtgcc aatgcagctg ctgctgatgg ccctggcatt tcacagcttg gcgaagtggc 3661 tcctctcccc accctgtctg ctcctgtgat ggagcccctc attaattctg agccttccac 3721 tggtgtgact gagccgctaa cgcctcatca gccaaaccac cagccctcct tcctcacctc 3781 tcctcaagac cttgcctctt ctccaatgga agaagatgag cagcattctg aagcagatga 3841 gcctccatca gacgaacccc tatctgatga ccccctgtca cctgctgagg agaaattgcc 3901 ccacattgat gagtattggt cagacagtga gcacatcttt ttggatgcaa atattggtgg 3961 ggtggccatc gcacctgctc acggctcggt tttgattgag tgtgcccggc gagagctgca 4021 cgctaccact cctgttgagc accccaaccg taatcatcca acccgcctct cccttgtctt 4081 ttaccagcac aaaaacctaa ataagcccca acatggtttt gaactaaaca agattaagtt 4141 tgaggctaaa gaagctaaga ataagaaaat gaaggcctca gagcaaaaag accaggcagc 4201 taatgaaggt ccagaacagt cctctgaagt aaatgaattg aaccaaattc cttctcataa 4261 agcattaaca ttaacccatg acaatgttgt caccgtgtcc ccttatgctc tcacacacgt 4321 tgcggggccc tataaccatt gggtctgatc tagagggccc ttcgaaggta agcctatccc 4381 taaccctctc ctcggtctcg attctacgcg taccggtcat catcaccatc accattgagt 4441 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc 4501 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa 4561 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg 4621 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg 4681 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc 4741 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact 4801 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc 4861 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt 4921 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc 4981 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt 5041 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat 5101 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa 5161 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc 5221 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc 5281 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg 5341 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat 5401 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc 5461 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct 5521 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa 5581 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac 5641 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg 5701 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag 5761 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt 5821 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg 5881 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg 5941 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga 6001 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag 6061 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat 6121 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt 6181 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg 6241 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt 6301 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc 6361 ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc 6421 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg 6481 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc 6541 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac 6601 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc 6661 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct 6721 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat 6781 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc 6841 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg 6901 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct 6961 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt 7021 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc aggaaccgta 7081 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa 7141 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc 7201 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt 7261 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca 7321 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg 7381 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat 7441 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta 7501 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct 7561 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac 7621 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa 7681 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa 7741 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt 7801 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca 7861 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca 7921 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc 7981 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa 8041 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc 8101 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca 8161 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat 8221 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag 8281 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac 8341 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt 8401 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt 8461 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc 8521 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat 8581 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca 8641 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga 8701 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg 8761 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg 8821 ttccgcgcac atttccccga aaagtgccac ctgacgt //