LOCUS       sequence-191771-        6045 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the human La antigen for use in ALARM NMR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6045)
  AUTHORS   Dahlin JL, Nelson KM, Strasser JM, Barsyte-Lovejoy D, Szewczyk MM, 
            Organ S, Cuellar M, Singh G, Shrimp JH, Nguyen N, Meier JL, 
            Arrowsmith CH, Brown PJ, Baell JB, Walters MA
  TITLE     Assay interference and off-target liabilities of reported histone 
            acetyltransferase inhibitors.
  JOURNAL   Nat Commun. 2017 Nov 15;8(1):1527. doi: 10.1038/s41467-017-01657-3.
  PUBMED    29142305
REFERENCE   2  (bases 1 to 6045)
  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: "Nat 
            Commun."; date: "2017-11-15"; volume: "8(1):1527. doi"; pages: " 
            10.1038/s41467-017-01657-3"
COMMENT     Sequence Label: ALARM NMR
FEATURES             Location/Qualifiers
     source          1..6045
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     69..87
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(140..157)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(857..877)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(887..919)
                     /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="MASMTGGQQMG"
     CDS             complement(923..940)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(950..967)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             986..1008
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1023..1047
                     /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)."
     primer_bind     complement(1047..1066)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1048..1066)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1150..1169)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1375..1452
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1453..2535
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(1472..1491)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3344..3535
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3529..3551)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3637..3776
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3692..3711
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3791..3808)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3962..4550)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4042..4061)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4661..4680)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4672..5487
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             4672..5487
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(4745..4764)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(5579..6034)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5716..5737)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5928..5947
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcttcttgtt
      181 ggtcttctat tattttcttc agtgcttctt tttccacctc tccttctagt acttcccaat
      241 tcacttcttt gttccttaat tgtaggttac cattatttgc atctttggct ttacccaatg
      301 cttccttggc tttttcttta aatagaatta tcccctcttt tgctcctctg acgaagtcta
      361 tccattttat ttcaccatga tttgaaaaaa gtatgtgtaa atcttctcta caggtctgat
      421 catctaaatc acccgaaaat ttcagcaagc atccaatctt ttcttctaga gatttcattt
      481 cagcatcttc ttctaacttt tgttttgctt cttgctcctg tttagctctt aatttagctt
      541 ccactttatt ttgttttctt tcttcatttt ttttggcaaa gtaatcgtcc ttgaaaagta
      601 ttagcaggtc tgtttctttg tacttctggc caggggtctc tacaaatttc ttagcagatt
      661 caatgctatc aaacacaaca aaaattgatc ccttaaatgc tttatgcaat gttcttctca
      721 tctgaatatt tagtacttga cctttatctt ctaaccattc ttttatgtca tcaagagttg
      781 catcagttgg gaagccttta atataaacag atctgttttt tacatcattt ttatactcat
      841 cagtcaccgg atccatgccc tgaaaataca ggttttccag atcccgaccc atttgctgtc
      901 caccagtcat gctagccata tggctgccgc gcggcaccag gccgctgctg tgatgatgat
      961 gatgatggct gctgcccatg gtatatctcc ttcttaaagt taaacaaaat tatttctaga
     1021 ggggaattgt tatccgctca caattcccct atagtgagtc gtattaattt cgcgggatcg
     1081 agatctcgat cctctacgcc ggacgcatcg tggccggcat caccggcgcc acaggtgcgg
     1141 ttgctggcgc ctatatcgcc gacatcaccg atggggaaga tcgggctcgc cacttcgggc
     1201 tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc cgtggccggg ggactgttgg
     1261 gcgccatctc cttgcatgca ccattccttg cggcggcggt gctcaacggc ctcaacctac
     1321 tactgggctg cttcctaatg caggagtcgc ataagggaga gcgtcgagat cccggacacc
     1381 atcgaatggc gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga gagtcaattc
     1441 agggtggtga atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc cggtgtctct
     1501 tatcagaccg tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa
     1561 aaagtggaag cggcgatggc ggagctgaat tacattccca accgcgtggc acaacaactg
     1621 gcgggcaaac agtcgttgct gattggcgtt gccacctcca gtctggccct gcacgcgccg
     1681 tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac tgggtgccag cgtggtggtg
     1741 tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg
     1801 caacgcgtca gtgggctgat cattaactat ccgctggatg accaggatgc cattgctgtg
     1861 gaagctgcct gcactaatgt tccggcgtta tttcttgatg tctctgacca gacacccatc
     1921 aacagtatta ttttctccca tgaagacggt acgcgactgg gcgtggagca tctggtcgca
     1981 ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg
     2041 cgtctggctg gctggcataa atatctcact cgcaatcaaa ttcagccgat agcggaacgg
     2101 gaaggcgact ggagtgccat gtccggtttt caacaaacca tgcaaatgct gaatgagggc
     2161 atcgttccca ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc
     2221 attaccgagt ccgggctgcg cgttggtgcg gatatctcgg tagtgggata cgacgatacc
     2281 gaagacagct catgttatat cccgccgtta accaccatca aacaggattt tcgcctgctg
     2341 gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt gaagggcaat
     2401 cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc
     2461 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     2521 gaaagcgggc agtgagcgca acgcaattaa tgtaagttag ctcactcatt aggcaccggg
     2581 atctcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg
     2641 catgactatc gtcgccgcac ttatgactgt cttctttatc atgcaactcg taggacaggt
     2701 gccggcagcg ctctgggtca ttttcggcga ggaccgcttt cgctggagcg cgacgatgat
     2761 cggcctgtcg cttgcggtat tcggaatctt gcacgccctc gctcaagcct tcgtcactgg
     2821 tcccgccacc aaacgtttcg gcgagaagca ggccattatc gccggcatgg cggccccacg
     2881 ggtgcgcatg atcgtgctcc tgtcgttgag gacccggcta ggctggcggg gttgccttac
     2941 tggttagcag aatgaatcac cgatacgcga gcgaacgtga agcgactgct gctgcaaaac
     3001 gtctgcgacc tgagcaacaa catgaatggt cttcggtttc cgtgtttcgt aaagtctgga
     3061 aacgcggaag tcagcgccct gcaccattat gttccggatc tgcatcgcag gatgctgctg
     3121 gctaccctgt ggaacaccta catctgtatt aacgaagcgc tggcattgac cctgagtgat
     3181 ttttctctgg tcccgccgca tccataccgc cagttgttta ccctcacaac gttccagtaa
     3241 ccgggcatgt tcatcatcag taacccgtat cgtgagcatc ctctctcgtt tcatcggtat
     3301 cattaccccc atgaacagaa atccccctta cacggaggca tcagtgacca aacaggaaaa
     3361 aaccgccctt aacatggccc gctttatcag aagccagaca ttaacgcttc tggagaaact
     3421 caacgagctg gacgcggatg aacaggcaga catctgtgaa tcgcttcacg accacgctga
     3481 tgagctttac cgcagctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     3541 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     3601 tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag
     3661 cgatagcgga gtgtatactg gcttaactat gcggcatcag agcagattgt actgagagtg
     3721 caccattgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgct
     3781 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     3841 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     3901 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     3961 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     4021 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     4081 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     4141 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     4201 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     4261 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     4321 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     4381 ctaactacgg ctacactaga aggaccgtat ttggtatctg cgctctgctg aagccagtta
     4441 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     4501 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     4561 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     4621 tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt tatgagccat
     4681 attcaacggg aaacgtcttg ctctaggccg cgattaaatt ccaacatgga tgctgattta
     4741 tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgattg
     4801 tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat
     4861 gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc
     4921 atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc gatccccggg
     4981 aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg
     5041 ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacagc
     5101 gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg
     5161 agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat
     5221 aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac
     5281 cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca
     5341 gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta
     5401 cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt
     5461 catttgatgc tcgatgagtt tttctaagaa ttaattcatg agcggataca tatttgaatg
     5521 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa
     5581 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     5641 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     5701 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     5761 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     5821 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     5881 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     5941 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     6001 cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt cgcca
//
