LOCUS       sequence-386978-        7666 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bacterial expression of sifA::HaloTag fusion, mutant form.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7666)
  TITLE     Hensel lab plasmids
REFERENCE   2  (bases 1 to 7666)
  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"
COMMENT     Sequence Label: p6187
FEATURES             Location/Qualifiers
     source          1..7666
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     348..367
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(586..690)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(716..1171)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(853..874)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1065..1084
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1298..1320
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1312..1329
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1313..1329
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1339..1358
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1339..1357
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2761..3645
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SGIARWLSTLEISG"
     CDS             3646..3672
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3718..3738)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3718..3736)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3757..3773)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3757..3773)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3770..3792)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3781..3797
                     /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(join(3805..3811,3812..3829,3830..3835))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3850..3871
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3988..4005)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      5150..5372
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             5420..6370
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     CDS             complement(join(7391..7666,1..516,517..585))
                     /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"
ORIGIN
        1 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
       61 ctcgtcgttt ggtatggctt cattcagctc cggttcccag cgatcaaggc gagttacatg
      121 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
      181 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
      241 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
      301 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
      361 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
      421 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
      481 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
      541 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
      601 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
      661 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt
      721 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
      781 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
      841 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
      901 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
      961 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     1021 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     1081 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     1141 gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct gcgcaactgt
     1201 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
     1261 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
     1321 acggccagtg agcgcgcgta atacgactca ctatagggcg aattgggtac caggtgagtg
     1381 atataagcga ttaattgcgc aacgctaaca aatccacacg catccaggca tgaagtttat
     1441 tcaagggtaa acttcatgcc ttcggcataa aaaacgcatg aaagaagttg ccgccagtat
     1501 tgcaaatcta caacatcatc cgcggtagtc cttcttttat ttttacctgt agcgacgcta
     1561 tcacagacag taatgcgttt atacgcgaag ctctcaggtt ttatactgat tgccagtctc
     1621 ttttaaaaat tatattacat ccgatgcgcc cgcagttgag ataaaaaggg tcgatttaat
     1681 caattatgta gtcattttta ctccagtata agtgagatta atatgccgat tactataggg
     1741 aatggttttt taaaaagtga aatccttacc aactccccaa ggaatacgaa agaagcatgg
     1801 tggaaagttt tatgggaaaa aattaaagac ttcttttttt ctactggcaa agcaaaagcg
     1861 gaccgttgtc tacatgagat gttgtttgcc gaacgcgccc ccacacgaga gcggcttaca
     1921 gagatttttt ttgagttgaa agagttagcc tgcgcatcgc aaagagatag atttcaggtt
     1981 cataatcctc atgaaaatga tgccaccatt attcttcgca tcatggatca aaacgaagag
     2041 aacgaattgt tacgtatcac tcaaaatacc gataccttta gctgtgaagt catggggaat
     2101 ctttattttt taatgaaaga tcgcccggat attttaaaat cgcatccaca aatgacggcc
     2161 atgattaaga gaagatatag cgaaatcgta gactaccccc tcccttcgac attatgtctc
     2221 aatcctgctg gcgcgccgat attatcggtt ccattagaca acatagaggg gtatttatat
     2281 actgaattga gaaaaggaca tttagatggg tggaaagcgc aagaaaaggc aacctacctg
     2341 gcagcgaaaa ttcagtctgg gattgaaaag acaacgcgca ttttacacca tgcgaatata
     2401 tccgaaagta ctcagcaaaa cgcattttta gaaacaatgg cgatgtgtgg attaaaacag
     2461 cttgaaatac caccaccgca tacccacata cctattgaaa aaatggtaaa agaggtttta
     2521 ctagcggata agacgtttca ggcgttcctc gtaacggatc ccagcaccag ccaaagtatg
     2581 ttagctgaga tagtcgaagc catctctgat caggtttttc acgccatttt tagaatagac
     2641 ccccaggcta tacaaaaaat ggcggaagaa cagttaacca cgctacacgt tcgctcagaa
     2701 caacaaagcg gcggctctgc ggcgtctgcg gcgggcgcgg gcgaagcggc ggcgggatcc
     2761 gaaatcggta ctggctttcc attcgacccc cattatgtgg aagtcctggg cgagcgcatg
     2821 cactacgtcg atgttggtcc gcgcgatggc acccctgtgc tgttcctgca cggtaacccg
     2881 acctcctcct acgtgtggcg caacatcatc ccgcatgttg caccgaccca tcgctgcatt
     2941 gctccagacc tgatcggtat gggcaaatcc gacaaaccag acctgggtta tttcttcgac
     3001 gaccacgtcc gcttcatgga tgccttcatc gaagccctgg gtctggaaga ggtcgtcctg
     3061 gtcattcacg actggggctc cgctctgggt ttccactggg ccaagcgcaa tccagagcgc
     3121 gtcaaaggta ttgcatttat ggagttcatc cgccctatcc cgacctggga cgaatggcca
     3181 gaatttgccc gcgagacctt ccaggccttc cgcaccaccg acgtcggccg caagctgatc
     3241 atcgatcaga acgtttttat cgagggtacg ctgccgatgg gtgtcgtccg cccgctgact
     3301 gaagtcgaga tggaccatta ccgcgagccg ttcctgaatc ctgttgaccg cgagccactg
     3361 tggcgcttcc caaacgagct gccaatcgcc ggtgagccag cgaacatcgt cgcgctggtc
     3421 gaagaataca tggactggct gcaccagtcc cctgtcccga agctgctgtt ctggggcacc
     3481 ccaggcgttc tgatcccacc ggccgaagcc gctcgcctgg ccaaaagcct gcctaactgc
     3541 aaggctgtgg acatcggccc gggtctgaat ctgctgcaag aagacaaccc ggacctgatc
     3601 ggcagcggga tcgcgcgctg gctgtctact ctggagattt ccggttaccc atacgacgtc
     3661 ccagactacg cttaatctag agcggccgcc accgcggtgg agctccagct tttgttccct
     3721 ttagtgaggg ttaattgcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     3781 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     3841 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     3901 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     3961 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     4021 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     4081 ggataacgca ggaaagaaca tgtgagatct ctacgggtcg gattttaagt cgtcttggta
     4141 ggaggcagcc tgaatggcga atgccgatgc ccttgagagc cttcaaccca gtcagctcct
     4201 tccggtgggc gcggggcatg actatcgtcg ccgcacttat gactgtcttc tttatcatgc
     4261 aactcgtagg acagggtgcc ggcagcgctc tgggtcattt tcggcgagga ccgctttcgc
     4321 tggagcgcga cgatgatcgg cctgtcgctt gcggtattcg gaatcttgca cgccctcgct
     4381 caagccttcg tcactggtcc cgccaccaaa cgtttcggcg agaagcaggc cattatcgcc
     4441 ggcatggcgg ccgacgcgct gggctacgtc ttgctggcgt tcgcgacgcg aggctggatg
     4501 gccttcccca ttatgattct tctcgcttcc ggcggcatcg ggatgcccgc gttgcaggcc
     4561 atgctgtcca ggcaggtaga tgacgaccat cagggacagc ttcaaggatc gctcgcggct
     4621 cttaccagcc taacttcgat cattggaccg ctgatcgtca cggcgattta tgccgcctcg
     4681 gcgagcacat ggaacgggtt ggcatggatt gtaggcgccg ccctatacct tgtctgcctc
     4741 cccgcgttgc gtcgcggtgc atggagccgg gccacctcga cctgaatgga agccggcggc
     4801 acctcgctaa cggattcacc actccgcaga cccgccataa aacgccctga gaagcccgtg
     4861 acgggctttt cttgtattat gggtagtttc cttgcatgaa tccataaaag gcgcctgtag
     4921 tgccatttac ccccattcac tgccagagcc gtgagcgcag cgaactgaat gtcacgaaaa
     4981 agacagcgac tcaggtgcct gatggtcgga gacaaaagga atattcagcg atttgcccga
     5041 gcttgcgagg gtgctactta atcctttagg gttttaaggt ctgttttgta gaggagcaaa
     5101 cagcgtttgc gacatccttt tgtaatactg cggaactgac taaagtagtg agttatacac
     5161 agggctggga tctattcttt ttatcttttt ttannntttc tttattctat aaattataac
     5221 cacttgaata taaacaaaaa aaacacacaa aggtctagcg gaatttacag agggtctagc
     5281 agaatttaca agttttccag caaaggtcta gcagaattta cagataccca caactcaaag
     5341 gaaaaggact agtaattatc attgactagc ccatctcaat tggtatagtg attaaaatca
     5401 cctagaccaa ttgagatgta tgtctgaatt agttgttttc aaagcaaatg aactagcgat
     5461 tagtcgctat gacttaacgg agcatgaaac caagctaatt ttatgctgtg tggcactact
     5521 caaccccacg attgaaaacc ctacaaggaa agaacggacg gtatcgttca cttataacca
     5581 atacgctcag atgatgaaca tcagtaggga aaatgcttat ggtgtattag ctaaagcaac
     5641 cagagagctg atgacgagaa ctgtggaaat caggaatcct ttggttaaag gctttgagat
     5701 tttccagtgg acaaactatg ccaagttctc aagcgaaaaa ttagaattag tttttagtga
     5761 agagatattg ccttatcttt tccagttaaa aaaattcata aaatataatc tggaacatgt
     5821 taagtctttt gaaaacaaat actctatgag gatttatgag tggttattaa aagaactaac
     5881 acaaaagaaa actcacaagg caaatataga gattagcctt gatgaattta agttcatgtt
     5941 aatgcttgaa aataactacc atgagtttaa aaggcttaac caatgggttt tgaaaccaat
     6001 aagtaaagat ttaaacactt acagcaatat gaaattggtg gttgataagc gaggccgccc
     6061 gactgatacg ttgattttcc aagttgaact agatagacaa atggatctcg taaccgaact
     6121 tgagaacaac cagataaaaa tgaatggtga caaaatacca acaaccatta catcagattc
     6181 ctacctacat aacggactaa gaaaaacact acacgatgct ttaactgcaa aaattcagct
     6241 caccagtttt gaggcaaaat ttttgagtga catgcaaagt aagtatgatc tcaatggttc
     6301 gttctcatgg ctcacgcaaa aacaacgaac cacactagag aacatactgg ctaaatacgg
     6361 aaggatctga ggttcttatg gctcttgtat ctatcagtga agcatcaaga ctaacaaaca
     6421 aaagtagaac aactgttcac cgttacatat caaagggaaa actgtccata tgcacagatg
     6481 aaaacggtgt aaaaaagata gatacatcag agcttttacg agtttttggt gcattcaaag
     6541 ctgttcacca tgaacagatc gacaatgtaa cagatgaaca gcatgtaaca cctaatagaa
     6601 caggtgaaac cagtaaaaca aagcaactag aacatgaaat tgaacacctg agacaacttg
     6661 ttacagctca acagtcacac atagacagcc tgaaacaggc gatgctgctt atcgaatcaa
     6721 agctgccgac aacacgggag ccagtgacgc ctcccgtggg gaaaaaatca tggcaattct
     6781 ggaagaaata gcgctttcag ccggcaaacc ggctgaagcc ggatctgcga ttctgataac
     6841 aaactagcaa caccagaaca gcccgtttgc gggcagcaaa acccgtactt ttggacgttc
     6901 cggcggtttt ttgtggcgag tggtgttcgg gcggtgcgcg caagatccat tatgttaaac
     6961 gggcgagttt acatctcaaa accgcccgct taacaccatc agaaatcctc agcgcgattt
     7021 taagcaccaa cccccccccg taacacccaa atccatactg aaagtggctt tgttgaataa
     7081 atcagatttc gggtaagtct cccccgtagc gggttgtgtt ttcaggcaat acgcacgctt
     7141 tcaggcatac ctgctttcgt cattttgttc agcgctcgta ccagggccat agcctccgca
     7201 acctgaccat cgtagtcacg cagcgtcagt gaacccccga acagagatct gacgctcagt
     7261 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     7321 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     7381 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     7441 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     7501 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     7561 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     7621 cctccatcca gtctattaat tgttgccggg aagctagagt aagtag
//
