LOCUS       sequence-437743-        5380 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpresses HTkC in E. coli cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5380)
  AUTHORS   Schwab S, Boyle AL, Dame RT
  TITLE     Novel histones and histone variant families in prokaryotes
  JOURNAL   bioRxiv 2023.06.01.543357
REFERENCE   2  (bases 1 to 5380)
  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: "bioRxiv 
            2023.06.01.543357"
COMMENT     Sequence Label: pRD503
FEATURES             Location/Qualifiers
     source          1..5380
                     /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"
     RBS             317..339
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    354..378
                     /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(378..397)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(379..397)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(485..504)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        710..787
                     /gene="lacI"
                     /label=lacI promoter
     CDS             788..1870
                     /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(807..826)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             2679..2870
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(2864..2886)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2972..3111
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3027..3046
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3126..3143)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3297..3885)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3377..3396)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(3996..4015)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4007..4822
                     /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             4007..4822
                     /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(4080..4099)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(4914..5369)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5051..5072)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5263..5282
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagt taaacatgac gtgcatacag ggttttacga ccttctgcct gtgcacgttt
      181 aattgctttt tcaaccaggg ctttaacttc tgcttccagt gcatcataaa attccggact
      241 aacacgcatt tccggatcaa cgcttttcac gaattctttc actttgctct taaccagcat
      301 ttctgccata tgtatatctc cttcttaaag ttaaacaaaa ttatttctag aggggaattg
      361 ttatccgctc acaattcccc tatagtgagt cgtattaatt tcgcgggatc gagatcgatc
      421 tcgatcctct acgccggacg catcgtggcc ggcatcaccg gcgccacagg tgcggttgct
      481 ggcgcctata tcgccgacat caccgatggg gaagatcggg ctcgccactt cgggctcatg
      541 agcgcttgtt tcggcgtggg tatggtggca ggccccgtgg ccgggggact gttgggcgcc
      601 atctccttgc atgcaccatt ccttgcggcg gcggtgctca acggcctcaa cctactactg
      661 ggctgcttcc taatgcagga gtcgcataag ggagagcgtc gagatcccgg acaccatcga
      721 atggcgcaaa acctttcgcg gtatggcatg atagcgcccg gaagagagtc aattcagggt
      781 ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag tatgccggtg tctcttatca
      841 gaccgtttcc cgcgtggtga accaggccag ccacgtttct gcgaaaacgc gggaaaaagt
      901 ggaagcggcg atggcggagc tgaattacat tcccaaccgc gtggcacaac aactggcggg
      961 caaacagtcg ttgctgattg gcgttgccac ctccagtctg gccctgcacg cgccgtcgca
     1021 aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt gccagcgtgg tggtgtcgat
     1081 ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg cacaatcttc tcgcgcaacg
     1141 cgtcagtggg ctgatcatta actatccgct ggatgaccag gatgccattg ctgtggaagc
     1201 tgcctgcact aatgttccgg cgttatttct tgatgtctct gaccagacac ccatcaacag
     1261 tattattttc tcccatgaag acggtacgcg actgggcgtg gagcatctgg tcgcattggg
     1321 tcaccagcaa atcgcgctgt tagcgggccc attaagttct gtctcggcgc gtctgcgtct
     1381 ggctggctgg cataaatatc tcactcgcaa tcaaattcag ccgatagcgg aacgggaagg
     1441 cgactggagt gccatgtccg gttttcaaca aaccatgcaa atgctgaatg agggcatcgt
     1501 tcccactgcg atgctggttg ccaacgatca gatggcgctg ggcgcaatgc gcgccattac
     1561 cgagtccggg ctgcgcgttg gtgcggacat ctcggtagtg ggatacgacg ataccgaaga
     1621 cagctcatgt tatatcccgc cgttaaccac catcaaacag gattttcgcc tgctggggca
     1681 aaccagcgtg gaccgcttgc tgcaactctc tcagggccag gcggtgaagg gcaatcagct
     1741 gttgcccgtc tcactggtga aaagaaaaac caccctggcg cccaatacgc aaaccgcctc
     1801 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     1861 cgggcagtga gcgcaacgca attaatgtaa gttagctcac tcattaggca ccgggatctc
     1921 gaccgatgcc cttgagagcc ttcaacccag tcagctcctt ccggtgggcg cggggcatga
     1981 ctatcgtcgc cgcacttatg actgtcttct ttatcatgca actcgtagga caggtgccgg
     2041 cagcgctctg ggtcattttc ggcgaggacc gctttcgctg gagcgcgacg atgatcggcc
     2101 tgtcgcttgc ggtattcgga atcttgcacg ccctcgctca agccttcgtc actggtcccg
     2161 ccaccaaacg tttcggcgag aagcaggcca ttatcgccgg catggcggcc ccacgggtgc
     2221 gcatgatcgt gctcctgtcg ttgaggaccc ggctaggctg gcggggttgc cttactggtt
     2281 agcagaatga atcaccgata cgcgagcgaa cgtgaagcga ctgctgctgc aaaacgtctg
     2341 cgacctgagc aacaacatga atggtcttcg gtttccgtgt ttcgtaaagt ctggaaacgc
     2401 ggaagtcagc gccctgcacc attatgttcc ggatctgcat cgcaggatgc tgctggctac
     2461 cctgtggaac acctacatct gtattaacga agcgctggca ttgaccctga gtgatttttc
     2521 tctggtcccg ccgcatccat accgccagtt gtttaccctc acaacgttcc agtaaccggg
     2581 catgttcatc atcagtaacc cgtatcgtga gcatcctctc tcgtttcatc ggtatcatta
     2641 cccccatgaa cagaaatccc ccttacacgg aggcatcagt gaccaaacag gaaaaaaccg
     2701 cccttaacat ggcccgcttt atcagaagcc agacattaac gcttctggag aaactcaacg
     2761 agctggacgc ggatgaacag gcagacatct gtgaatcgct tcacgaccac gctgatgagc
     2821 tttaccgcag ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     2881 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     2941 gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat gacccagtca cgtagcgata
     3001 gcggagtgta tactggctta actatgcggc atcagagcag attgtactga gagtgcacca
     3061 ttgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gcgctcttcc
     3121 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     3181 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     3241 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     3301 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     3361 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     3421 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     3481 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     3541 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     3601 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     3661 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     3721 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     3781 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     3841 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ttcaagaaga tcctttgatc
     3901 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     3961 aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca
     4021 acgggaaacg tcttgctcta ggccgcgatt aaattccaac atggatgctg atttatatgg
     4081 gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc gattgtatgg
     4141 gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt
     4201 tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa
     4261 gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc ccgggaaaac
     4321 agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc
     4381 agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg
     4441 cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga
     4501 ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact
     4561 tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat
     4621 ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg
     4681 ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa
     4741 acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt
     4801 gatgctcgat gagtttttct aagaattaat tcatgagcgg atacatattt gaatgtattt
     4861 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctaaattgt
     4921 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     4981 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     5041 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     5101 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     5161 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     5221 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     5281 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     5341 cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca
//
