LOCUS       sequence-388083-        7684 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bacterial expression of sifA::HaloTag fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7684)
  TITLE     Hensel lab plasmids
REFERENCE   2  (bases 1 to 7684)
  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: p4305
FEATURES             Location/Qualifiers
     source          1..7684
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1297..2247)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      2295..2517
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     primer_bind     3662..3679
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3796..3817
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3832..3837,3838..3855,3856..3862)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3870..3886
                     /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     3875..3897
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3894..3910
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3894..3910
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3929..3949
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3931..3949
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             complement(3995..4021)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(4022..4906)
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SGIARWLSTLEISG"
     primer_bind     complement(6327..6346)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6328..6346)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6356..6373)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6356..6372)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6365..6387)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6514..6969
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6601..6620)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6811..6832
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6995..7099
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7100..7168,7169..7684,1..276)
                     /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"
     primer_bind     complement(7318..7337)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
       61 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
      121 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
      181 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
      241 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
      301 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
      361 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagat
      421 ctctgttcgg gggttcactg acgctgcgtg actacgatgg tcaggttgcg gaggctatgg
      481 ccctggtacg agcgctgaac aaaatgacga aagcaggtat gcctgaaagc gtgcgtattg
      541 cctgaaaaca caacccgcta cgggggagac ttacccgaaa tctgatttat tcaacaaagc
      601 cactttcagt atggatttgg gtgttacggg ggggggttgg tgcttaaaat cgcgctgagg
      661 atttctgatg gtgttaagcg ggcggttttg agatgtaaac tcgcccgttt aacataatgg
      721 atcttgcgcg caccgcccga acaccactcg ccacaaaaaa ccgccggaac gtccaaaagt
      781 acgggttttg ctgcccgcaa acgggctgtt ctggtgttgc tagtttgtta tcagaatcgc
      841 agatccggct tcagccggtt tgccggctga aagcgctatt tcttccagaa ttgccatgat
      901 tttttcccca cgggaggcgt cactggctcc cgtgttgtcg gcagctttga ttcgataagc
      961 agcatcgcct gtttcaggct gtctatgtgt gactgttgag ctgtaacaag ttgtctcagg
     1021 tgttcaattt catgttctag ttgctttgtt ttactggttt cacctgttct attaggtgtt
     1081 acatgctgtt catctgttac attgtcgatc tgttcatggt gaacagcttt gaatgcacca
     1141 aaaactcgta aaagctctga tgtatctatc ttttttacac cgttttcatc tgtgcatatg
     1201 gacagttttc cctttgatat gtaacggtga acagttgttc tacttttgtt tgttagtctt
     1261 gatgcttcac tgatagatac aagagccata agaacctcag atccttccgt atttagccag
     1321 tatgttctct agtgtggttc gttgtttttg cgtgagccat gagaacgaac cattgagatc
     1381 atacttactt tgcatgtcac tcaaaaattt tgcctcaaaa ctggtgagct gaatttttgc
     1441 agttaaagca tcgtgtagtg tttttcttag tccgttatgt aggtaggaat ctgatgtaat
     1501 ggttgttggt attttgtcac cattcatttt tatctggttg ttctcaagtt cggttacgag
     1561 atccatttgt ctatctagtt caacttggaa aatcaacgta tcagtcgggc ggcctcgctt
     1621 atcaaccacc aatttcatat tgctgtaagt gtttaaatct ttacttattg gtttcaaaac
     1681 ccattggtta agccttttaa actcatggta gttattttca agcattaaca tgaacttaaa
     1741 ttcatcaagg ctaatctcta tatttgcctt gtgagttttc ttttgtgtta gttcttttaa
     1801 taaccactca taaatcctca tagagtattt gttttcaaaa gacttaacat gttccagatt
     1861 atattttatg aattttttta actggaaaag ataaggcaat atctcttcac taaaaactaa
     1921 ttctaatttt tcgcttgaga acttggcata gtttgtccac tggaaaatct caaagccttt
     1981 aaccaaagga ttcctgattt ccacagttct cgtcatcagc tctctggttg ctttagctaa
     2041 tacaccataa gcattttccc tactgatgtt catcatctga gcgtattggt tataagtgaa
     2101 cgataccgtc cgttctttcc ttgtagggtt ttcaatcgtg gggttgagta gtgccacaca
     2161 gcataaaatt agcttggttt catgctccgt taagtcatag cgactaatcg ctagttcatt
     2221 tgctttgaaa acaactaatt cagacataca tctcaattgg tctaggtgat tttaatcact
     2281 ataccaattg agatgggcta gtcaatgata attactagtc cttttccttt gagttgtggg
     2341 tatctgtaaa ttctgctaga cctttgctgg aaaacttgta aattctgcta gaccctctgt
     2401 aaattccgct agacctttgt gtgttttttt tgtttatatt caagtggtta taatttatag
     2461 aataaagaaa gaataaaaaa agataaaaag aatagatccc agccctgtgt ataactcact
     2521 actttagtca gttccgcagt attacaaaag gatgtcgcaa acgctgtttg ctcctctaca
     2581 aaacagacct taaaacccta aaggattaag tagcaccctc gcaagctcgg gcaaatcgct
     2641 gaatattcct tttgtctccg accatcaggc acctgagtcg ctgtcttttt cgtgacattc
     2701 agttcgctgc gctcacggct ctggcagtga atgggggtaa atggcactac aggcgccttt
     2761 tatggattca tgcaaggaaa ctacccataa tacaagaaaa gcccgtcacg ggcttctcag
     2821 ggcgttttat ggcgggtctg cggagtggtg aatccgttag cgaggtgccg ccggcttcca
     2881 ttcaggtcga ggtggcccgg ctccatgcac cgcgacgcaa cgcggggagg cagacaaggt
     2941 atagggcggc gcctacaatc catgccaacc cgttccatgt gctcgccgag gcggcataaa
     3001 tcgccgtgac gatcagcggt ccaatgatcg aagttaggct ggtaagagcc gcgagcgatc
     3061 cttgaagctg tccctgatgg tcgtcatcta cctgcctgga cagcatggcc tgcaacgcgg
     3121 gcatcccgat gccgccggaa gcgagaagaa tcataatggg gaaggccatc cagcctcgcg
     3181 tcgcgaacgc cagcaagacg tagcccagcg cgtcggccgc catgccggcg ataatggcct
     3241 gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca
     3301 agattccgaa taccgcaagc gacaggccga tcatcgtcgc gctccagcga aagcggtcct
     3361 cgccgaaaat gacccagagc gctgccggca ccctgtccta cgagttgcat gataaagaag
     3421 acagtcataa gtgcggcgac gatagtcatg ccccgcgccc accggaagga gctgactggg
     3481 ttgaaggctc tcaagggcat cggcattcgc cattcaggct gcctcctacc aagacgactt
     3541 aaaatccgac ccgtagagat ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     3601 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     3661 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     3721 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     3781 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     3841 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     3901 cagctatgac catgattacg ccaagcgcgc aattaaccct cactaaaggg aacaaaagct
     3961 ggagctccac cgcggtggcg gccgctctag attaagcgta gtctgggacg tcgtatgggt
     4021 aaccggaaat ctccagagta gacagccagc gcgcgatccc gctgccgatc aggtccgggt
     4081 tgtcttcttg cagcagattc agacccgggc cgatgtccac agccttgcag ttaggcaggc
     4141 ttttggccag gcgagcggct tcggccggtg ggatcagaac gcctggggtg ccccagaaca
     4201 gcagcttcgg gacaggggac tggtgcagcc agtccatgta ttcttcgacc agcgcgacga
     4261 tgttcgctgg ctcaccggcg attggcagct cgtttgggaa gcgccacagt ggctcgcggt
     4321 caacaggatt caggaacggc tcgcggtaat ggtccatctc gacttcagtc agcgggcgga
     4381 cgacacccat cggcagcgta ccctcgataa aaacgttctg atcgatgatc agcttgcggc
     4441 cgacgtcggt ggtgcggaag gcctggaagg tctcgcgggc aaattctggc cattcgtccc
     4501 aggtcgggat agggcggatg aactccataa atgcaatacc tttgacgcgc tctggattgc
     4561 gcttggccca gtggaaaccc agagcggagc cccagtcgtg aatgaccagg acgacctctt
     4621 ccagacccag ggcttcgatg aaggcatcca tgaagcggac gtggtcgtcg aagaaataac
     4681 ccaggtctgg tttgtcggat ttgcccatac cgatcaggtc tggagcaatg cagcgatggg
     4741 tcggtgcaac atgcgggatg atgttgcgcc acacgtagga ggaggtcggg ttaccgtgca
     4801 ggaacagcac aggggtgcca tcgcgcggac caacatcgac gtagtgcatg cgctcgccca
     4861 ggacttccac ataatggggg tcgaatggaa agccagtacc gatttcggat cccgccgccg
     4921 cttcgcccgc gcccgccgca gacgccgcag agcctaaaaa acaacataaa cagccgcttt
     4981 gttgttctga gcgaacgtgt agcgtggtta actgttcttc cgccattttt tgtatagcct
     5041 gggggtctat tctaaaaatg gcgtgaaaaa cctgatcaga gatggcttcg actatctcag
     5101 ctaacatact ttggctggtg ctgggatccg ttacgaggaa cgcctgaaac gtcttatccg
     5161 ctagtaaaac ctcttttacc attttttcaa taggtatgtg ggtatgcggt ggtggtattt
     5221 caagctgttt taatccacac atcgccattg tttctaaaaa tgcgttttgc tgagtacttt
     5281 cggatatatt cgcatggtgt aaaatgcgcg ttgtcttttc aatcccagac tgaattttcg
     5341 ctgccaggta ggttgccttt tcttgcgctt tccacccatc taaatgtcct tttctcaatt
     5401 cagtatataa atacccctct atgttgtcta atggaaccga taatatcggc gcgccagcag
     5461 gattgagaca taatgtcgaa gggagggggt agtctacgat ttcgctatat cttctcttaa
     5521 tcatggccgt catttgtgga tgcgatttta aaatatccgg gcgatctttc attaaaaaat
     5581 aaagattccc catgacttca cagctaaagg tatcggtatt ttgagtgata cgtaacaatt
     5641 cgttctcttc gttttgatcc atgatgcgaa gaataatggt ggcatcattt tcatgaggat
     5701 tatgaacctg aaatctatct ctttgcgatg cgcaggctaa ctctttcaac tcaaaaaaaa
     5761 tctctgtaag ccgctctcgt gtgggggcgc gttcggcaaa caacatctca tgtagacaac
     5821 ggtccgcttt tgctttgcca gtagaaaaaa agaagtcttt aattttttcc cataaaactt
     5881 tccaccatgc ttctttcgta ttccttgggg agttggtaag gatttcactt tttaaaaaac
     5941 cattccctat agtaatcggc atattaatct cacttatact ggagtaaaaa tgactacata
     6001 attgattaaa tcgacccttt ttatctcaac tgcgggcgca tcggatgtaa tataattttt
     6061 aaaagagact ggcaatcagt ataaaacctg agagcttcgc gtataaacgc attactgtct
     6121 gtgatagcgt cgctacaggt aaaaataaaa gaaggactac cgcggatgat gttgtagatt
     6181 tgcaatactg gcggcaactt ctttcatgcg ttttttatgc cgaaggcatg aagtttaccc
     6241 ttgaataaac ttcatgcctg gatgcgtgtg gatttgttag cgttgcgcaa ttaatcgctt
     6301 atatcactca cctggtaccc aattcgccct atagtgagtc gtattacgcg cgctcactgg
     6361 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
     6421 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
     6481 cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg
     6541 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     6601 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     6661 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     6721 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     6781 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     6841 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     6901 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
     6961 ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     7021 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     7081 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     7141 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     7201 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     7261 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     7321 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     7381 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     7441 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     7501 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     7561 ggatcatgta actcgccttg atcgctggga accggagctg aatgaagcca taccaaacga
     7621 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     7681 cgaa
//
