LOCUS       sequence-393193-        3814 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Bacterial expression plasmid with Halotag v7 insert plus a 
            C-terminus appended AviTag for enzymatic biotinylation..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3814)
  TITLE     Bacterial expression plasmids of HaloTag v7 variants
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3814)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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            "Unpublished"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.AviTag
FEATURES             Location/Qualifiers
     source          1..3814
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     25..44
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        25..43
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             75..97
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             117..134
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             138..170
                     /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(174..182,183..197)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             213..1103
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     CDS             1110..1154
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(1220..1238)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1234..1281
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1378..1833
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1465..1484)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1675..1696
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1859..1963
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1964..2032,2033..2824)
                     /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(2182..2201)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2995..3583
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3484..3503
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3737..3754
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tgcaggatct cgatcccgcg aaattaatac gactcactat agggagacca caacggtttc
       61 cctctagaaa taattttgtt taactttaag aaggagatat acatatgcgg ggttctcatc
      121 atcatcatca tcatggtatg gctagcatga ctggtggaca gcaaatgggt cgggatctgt
      181 acgacgatga cgataaggat cgatggggat ccatggcaga aatcggtact ggctttccat
      241 tcgaccccca ttatgtggaa gtcctgggcg agcgcatgca ctacgtcgat gttggtccgc
      301 gcgatggcac ccctgtgctg ttcctgcacg gtaacccgac ctcctcctac gtgtggcgca
      361 acatcatccc gcatgttgca ccgacccatc gctgcattgc tccagacctg atcggtatgg
      421 gcaaatccga caaaccagac ctgggttatt tcttcgacga ccacgtccgc ttcatggatg
      481 ccttcatcga agccctgggt ctggaagagg tcgtcctggt cattcacgac tggggctccg
      541 ctctgggttt ccactgggcc aagcgcaatc cagagcgcgt caaaggtatt gcatttatgg
      601 agttcatccg ccctatcccg acctgggacg aatggccaga atttgcccgc gagaccttcc
      661 aggccttccg caccaccgac gtcggccgca agctgatcat cgatcagaac gtttttatcg
      721 agggtacgct gccgatgggt gtcgtccgcc cgctgactga agtcgagatg gaccattacc
      781 gcgagccgtt cctgaatcct gttgaccgcg agccactgtg gcgcttccca aacgagctgc
      841 caatcgccgg tgagccagcg aacatcgtcg cgctggtcga agaatacatg gactggctgc
      901 accagtcccc tgtcccgaag ctgctgttct ggggcacccc aggcgttctg atcccaccgg
      961 ccgaagccgc tcgcctggcc aaaagcctgc ctaactgcaa ggctgtggac atcggcccgg
     1021 gtctgaatct gctgcaagaa gacaaccccg acctgatcgg cagcgagatc gcgcgctggc
     1081 tgtcgacgct cgagatttcc ggtgggtccg ggttgaatga tattttcgaa gcacagaaaa
     1141 ttgaatggca tgagtaagaa ttcgaagctt gatccggctg ctaacaaagc ccgaaaggaa
     1201 gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa
     1261 cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggatctgg cgtaatagcg
     1321 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgggacg
     1381 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     1441 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     1501 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     1561 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     1621 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     1681 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     1741 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     1801 cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg
     1861 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     1921 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     1981 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     2041 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     2101 actggatctc aacagcggta agatccttga gagttttcgc cccaaagaac gttttccaat
     2161 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     2221 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     2281 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     2341 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     2401 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     2461 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     2521 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     2581 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     2641 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     2701 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     2761 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     2821 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     2881 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     2941 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     3001 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     3061 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     3121 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     3181 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     3241 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     3301 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     3361 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     3421 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     3481 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     3541 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     3601 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     3661 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     3721 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     3781 accgcctctc cccgcgcgtt ggccgattca ttaa
//
