LOCUS       sequence-393190-        3763 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid with HaloTag v7 insert; modified to 
            replace cysteines C61, C262, and optimized to human codon 
            bias/preference for use in oxidizing environments..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3763)
  TITLE     Bacterial expression plasmids of HaloTag v7 variants
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3763)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pRSET.HaloTag.v7.CoOp.Cys-less
FEATURES             Location/Qualifiers
     source          1..3763
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     20..39
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        20..38
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             70..92
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             112..129
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             133..165
                     /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             join(169..177,178..192)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     primer_bind     complement(1164..1182)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1178..1225
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1322..1777
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1409..1428)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1619..1640
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1803..1907
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1908..1976,1977..2768)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPKERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2126..2145)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2939..3527
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3428..3447
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3681..3698
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
       61 agaaataatt ttgtttaact ttaagaagga gatatacata tgcggggttc tcatcatcat
      121 catcatcatg gtatggctag catgactggt ggacagcaaa tgggtcggga tctgtacgac
      181 gatgacgata aggatcgatg gggatccatg gccgagatcg gcaccggctt ccccttcgac
      241 ccccactacg tggaggtgct gggcgagcgc atgcactacg ttgacgtggg cccccgcgac
      301 ggcacccccg tgctgttcct gcacggcaac cccaccagca gctacgtgtg gcgcaacatc
      361 atcccccacg tggcccccac ccaccgcact atcgcccccg acctgatcgg catgggcaag
      421 agcgacaagc ccgacctggg ctacttcttc gacgaccacg tgcgcttcat ggacgccttc
      481 atcgaggccc tgggcctgga ggaggtggtg ctggtgatcc acgactgggg cagcgccctg
      541 ggcttccact gggccaagcg caaccccgag cgcgtgaagg gcatcgcctt catggagttc
      601 atccgcccca tccccacctg ggacgagtgg cccgagttcg cccgcgagac cttccaggcc
      661 ttccgcacca ccgacgtggg ccgcaagctg atcatcgacc agaacgtgtt catcgagggc
      721 accctgccca tgggcgtggt gcgccccctg accgaggtgg agatggacca ctaccgcgag
      781 cccttcctga accccgtgga ccgcgagccc ctgtggcgct tccccaacga gctgcccatc
      841 gccggcgagc ccgccaacat cgtggccctg gtggaggagt acatggactg gctgcaccag
      901 agccccgtgc ccaagctgct gttctggggc acccccggcg tgctgatccc ccccgccgag
      961 gccgcccgcc tggccaagag cctgcccaac gttaaggccg tggacatcgg ccccggcctg
     1021 aacctgctcc aggaggacaa ccccgacctg atcggcagcg agatcgcccg ctggctgagc
     1081 accctggaga tcagcggctg agaattcgaa gcttgatccg gctgctaaca aagcccgaaa
     1141 ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc
     1201 taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggat ctggcgtaat
     1261 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     1321 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     1381 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     1441 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     1501 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     1561 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     1621 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     1681 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     1741 aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg
     1801 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     1861 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     1921 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     1981 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     2041 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccaaa gaacgttttc
     2101 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     2161 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     2221 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     2281 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     2341 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     2401 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     2461 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     2521 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     2581 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     2641 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     2701 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     2761 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     2821 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     2881 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     2941 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     3001 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     3061 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     3121 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     3181 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     3241 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     3301 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     3361 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     3421 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     3481 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     3541 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     3601 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     3661 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     3721 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cag
//
