LOCUS       sequence-388057-        7775 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bacterial expression of pipB2::HaloTag fusion, mutant form .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7775)
  TITLE     Hensel lab plasmids
REFERENCE   2  (bases 1 to 7775)
  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: p6172
FEATURES             Location/Qualifiers
     source          1..7775
                     /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(6418..6437)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6419..6437)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6447..6464)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6447..6463)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6456..6478)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6605..7060
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6692..6711)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6902..6923
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7086..7190
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7191..7259,7260..7775,1..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(7409..7428)
                     /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 agccaatatt ttcactataa aattcgttaa
     4981 agagtgtttg tgtgcttgta gacattgtgg cgtcttcagt acgcaaatcg gttagtgtac
     5041 aatctttaaa acaggtattg tttaaattag cctcagccaa atcagccccg tccaggttcg
     5101 tagaaatgaa actcgccttt cgcaggtttg cccgggttaa gacagcgcct tccataattg
     5161 cagccattat agtagcaccc tgtaatgtcg caccgctcat atcagcacgt ataagggttg
     5221 cgcctgaaag attagtgtga tccatatttg caccggagca attacattct atcatgctac
     5281 agcctaacag actcgcgtgt tgtaaattgg cgccagtaag attagcgtga tcgagggtag
     5341 cgccacacat aatgctatct tccagacagg cgttcttgaa attactgtcg cctagtgatg
     5401 ttcccgatag atttgcgccc ttaaaatttg caccttcaca atccgccata aacagcactg
     5461 ccccctctaa atttgcgccg cttaagtctg cgtcacataa attttctgac gtcagatttg
     5521 ctccggtaag gacagcgcct ttaagattca tgcctccctg ggcagttaat attacagaat
     5581 cctgagggag agaaaattcg catctgaata gtaaagctcg gcatattttt tcaaaagagg
     5641 caagcggaat ttctcttgtc tcagaatggt cgcctttact gactcgtaca ataacttgtc
     5701 cagcttcgtt attctcacca ggaagattaa attcgacacg acacccatcc atatcatcca
     5761 ggatgatgtt ttctggcaac ggagcacctc tgtcaggcat tgtacttttc aatgtctcag
     5821 catcagtctc tatgtcttcc tgatcttgtg tttcatttct ccgacgtatc ccaccacagg
     5881 taaaaaagtt aatgatatat tccagaatgg ttttgggcga ggtagcttgc cgcatagcag
     5941 cagaagtccc cgcgccaaaa gcagatttag ccataccagc cagactatcg agtgaacgct
     6001 ccatatattt tctcccagag acagcaacac agtgcatgat aaaatttatc atatagattt
     6061 gcatttagag ataaggtgaa tacccccttt acgtctggtt acataataac gtatcacgac
     6121 gcgttgctgt ctggcttaca caatagcagc caaataaata tattggctta tactcgtcat
     6181 acttcaagtt gcatgtgctg cggccgcgtt ccctcacccc agtcacttac tttagtaagc
     6241 tcctggggat tcgctcgctt gccgccttcc tgcaactcga attatttaga gtatataaca
     6301 tcatttaaaa taataaacac ggcattcgtt ttaatactac gttctggact catcccactc
     6361 attagcagaa aacctattaa aaagcgtgtt caggcatttt attaggtacc caattcgccc
     6421 tatagtgagt cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa
     6481 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     6541 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     6601 tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     6661 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     6721 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     6781 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     6841 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     6901 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     6961 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     7021 tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa
     7081 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     7141 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     7201 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     7261 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     7321 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     7381 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     7441 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     7501 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     7561 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     7621 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgctggg
     7681 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     7741 tggcaacaac gttgcgcaaa ctattaactg gcgaa
//
