LOCUS       sequence-387011-        7682 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bacterial expression of sopD2::HaloTag fusion, mutant form.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7682)
  TITLE     Hensel lab plasmids
REFERENCE   2  (bases 1 to 7682)
  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: p6173
FEATURES             Location/Qualifiers
     source          1..7682
                     /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             2777..3661
                     /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             3662..3688
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3734..3754)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3734..3752)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3773..3789)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3773..3789)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3786..3808)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3797..3813
                     /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(3821..3827,3828..3845,3846..3851))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3866..3887
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4004..4021)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      5166..5388
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             5436..6386
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLDNILAKYGRI"
     CDS             complement(join(7407..7682,1..516,517..585))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDSGQEQLGRRIHYSQNDLVEYS
                     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 cccggagtca 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 tgatacatat 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 ccagtataaa
     1381 atatacggca aatctatgct gcataactcg caccgccaga aattctggct cattcatcag
     1441 ctcgtaacgc ctcaaccgca atacatactg caagtgtatc atccatagcg ccctggtttt
     1501 actgtttatg atccgcctct tttttcgaat ggttggcgcc gttttgttgt ggtcagaacg
     1561 aaaataattc tcaacataat tcagatgtgt ccaaagaacg ttatgcgctg tccagcgcct
     1621 ggcgtcgcat tcaacgcgca atcaggtgta aatctgatgt catttctaaa cccaggctga
     1681 ttcaatctct taaatagagt gtggttttaa tcaaaaaatg agagcaacgg attggatctt
     1741 gctttcgcgg taaataatca agggagttat tatgccagtt acgttaagtt ttggtaatcg
     1801 tcataactat gaaattaatc attcacggct agcccgtttg atgagtcctg ataaagaaga
     1861 agcgctatac atgggggtat gggatagatt taaggactgc ttcagaacac acaaaaaaca
     1921 agaggtgctg gaggtattat atacactcat ccatggatgt gaacgtgaaa atcaagctga
     1981 acttaatgtt gatataaccg gcatggagaa gatacatgcc tttacacaat taaaagaata
     2041 tgctaatcca tctcaacaag accgctttgt tatgcgtttt gatatgaatc agacacaagt
     2101 tttatttgag attgatggca aggttattga taaatgtaat ttgcatcgtc tcctcaatgt
     2161 ttcagaaaac tgtattttta aagtgatgga agaagatgaa gaagagttgt ttcttaaaat
     2221 atgtattaag tatggagaaa aaatttctcg ctatccagaa ttattagaag gatttgctaa
     2281 taaacttaaa gatgcagtta atgaggatga tgacgttaag gatgaagttt ataagcttat
     2341 gcgatctgga gaagatcgaa aaatggaatg tgttgaatgg aatggtactc ttactgaaga
     2401 ggagaagaat aaactacgct gtcttcaaat ggggtctttt aatattacta cccaattttt
     2461 taaaataggt tactgggaat tagaggggga agtcctgttt gatatggttc atccaacact
     2521 tagctatctg ctacaagcat ataaaccgtc attgtcatct gatttaatag aaactaacac
     2581 catgctgttt tctgatgtat taaataagga tgatgatgag tctcaaaata atgacagaga
     2641 aattgatgct attctaagaa gaatatatag atcacataat aacactttat ttataagtga
     2701 aaagtcgagt tgtcgcaata tgcttatagg ctctgcggcg tctgcggcgg gcgcgggcga
     2761 agcggcggcg ggatccgaaa tcggtactgg ctttccattc gacccccatt atgtggaagt
     2821 cctgggcgag cgcatgcact acgtcgatgt tggtccgcgc gatggcaccc ctgtgctgtt
     2881 cctgcacggt aacccgacct cctcctacgt gtggcgcaac atcatcccgc atgttgcacc
     2941 gacccatcgc tgcattgctc cagacctgat cggtatgggc aaatccgaca aaccagacct
     3001 gggttatttc ttcgacgacc acgtccgctt catggatgcc ttcatcgaag ccctgggtct
     3061 ggaagaggtc gtcctggtca ttcacgactg gggctccgct ctgggtttcc actgggccaa
     3121 gcgcaatcca gagcgcgtca aaggtattgc atttatggag ttcatccgcc ctatcccgac
     3181 ctgggacgaa tggccagaat ttgcccgcga gaccttccag gccttccgca ccaccgacgt
     3241 cggccgcaag ctgatcatcg atcagaacgt ttttatcgag ggtacgctgc cgatgggtgt
     3301 cgtccgcccg ctgactgaag tcgagatgga ccattaccgc gagccgttcc tgaatcctgt
     3361 tgaccgcgag ccactgtggc gcttcccaaa cgagctgcca atcgccggtg agccagcgaa
     3421 catcgtcgcg ctggtcgaag aatacatgga ctggctgcac cagtcccctg tcccgaagct
     3481 gctgttctgg ggcaccccag gcgttctgat cccaccggcc gaagccgctc gcctggccaa
     3541 aagcctgcct aactgcaagg ctgtggacat cggcccgggt ctgaatctgc tgcaagaaga
     3601 caacccggac ctgatcggca gcgggatcgc gcgctggctg tctactctgg agatttccgg
     3661 ttacccatac gacgtcccag actacgctta atctagagcg gccgccaccg cggtggagct
     3721 ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc
     3781 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     3841 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     3901 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     3961 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     4021 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     4081 tatccacaga atcaggggat aacgcaggaa agaacatgtg agatctctac gggtcggatt
     4141 ttaagtcgtc ttggtaggag gcagcctgaa tggcgaatgc cgatgccctt gagagccttc
     4201 aacccagtca gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc acttatgact
     4261 gtcttcttta tcatgcaact cgtaggacag ggtgccggca gcgctctggg tcattttcgg
     4321 cgaggaccgc tttcgctgga gcgcgacgat gatcggcctg tcgcttgcgg tattcggaat
     4381 cttgcacgcc ctcgctcaag ccttcgtcac tggtcccgcc accaaacgtt tcggcgagaa
     4441 gcaggccatt atcgccggca tggcggccga cgcgctgggc tacgtcttgc tggcgttcgc
     4501 gacgcgaggc tggatggcct tccccattat gattcttctc gcttccggcg gcatcgggat
     4561 gcccgcgttg caggccatgc tgtccaggca ggtagatgac gaccatcagg gacagcttca
     4621 aggatcgctc gcggctctta ccagcctaac ttcgatcatt ggaccgctga tcgtcacggc
     4681 gatttatgcc gcctcggcga gcacatggaa cgggttggca tggattgtag gcgccgccct
     4741 ataccttgtc tgcctccccg cgttgcgtcg cggtgcatgg agccgggcca cctcgacctg
     4801 aatggaagcc ggcggcacct cgctaacgga ttcaccactc cgcagacccg ccataaaacg
     4861 ccctgagaag cccgtgacgg gcttttcttg tattatgggt agtttccttg catgaatcca
     4921 taaaaggcgc ctgtagtgcc atttaccccc attcactgcc agagccgtga gcgcagcgaa
     4981 ctgaatgtca cgaaaaagac agcgactcag gtgcctgatg gtcggagaca aaaggaatat
     5041 tcagcgattt gcccgagctt gcgagggtgc tacttaatcc tttagggttt taaggtctgt
     5101 tttgtagagg agcaaacagc gtttgcgaca tccttttgta atactgcgga actgactaaa
     5161 gtagtgagtt atacacaggg ctgggatcta ttctttttat ctttttttat tctttcttta
     5221 ttctataaat tataaccact tgaatataaa caaaaaaaac acacaaaggt ctagcggaat
     5281 ttacagaggg tctagcagaa tttacaagtt ttccagcaaa ggtctagcag aatttacaga
     5341 tacccacaac tcaaaggaaa aggactagta attatcattg actagcccat ctcaattggt
     5401 atagtgatta aaatcaccta gaccaattga gatgtatgtc tgaattagtt gttttcaaag
     5461 caaatgaact agcgattagt cgctatgact taacggagca tgaaaccaag ctaattttat
     5521 gctgtgtggc actactcaac cccacgattg aaaaccctac aaggaaagaa cggacggtat
     5581 cgttcactta taaccaatac gctcagatga tgaacatcag tagggaaaat gcttatggtg
     5641 tattagctaa agcaaccaga gagctgatga cgagaactgt ggaaatcagg aatcctttgg
     5701 ttaaaggctt tgagattttc cagtggacaa actatgccaa gttctcaagc gaaaaattag
     5761 aattagtttt tagtgaagag atattgcctt atcttttcca gttaaaaaaa ttcataaaat
     5821 ataatctgga acatgttaag tcttttgaaa acaaatactc tatgaggatt tatgagtggt
     5881 tattaaaaga actaacacaa aagaaaactc acaaggcaaa tatagagatt agccttgatg
     5941 aatttaagtt catgttaatg cttgaaaata actaccatga gtttaaaagg cttaaccaat
     6001 gggttttgaa accaataagt aaagatttaa acacttacag caatatgaaa ttggtggttg
     6061 ataagcgagg ccgcccgact gatacgttga ttttccaagt tgaactagat agacaaatgg
     6121 atctcgtaac cgaacttgag aacaaccaga taaaaatgaa tggtgacaaa ataccaacaa
     6181 ccattacatc agattcctac ctacataacg gactaagaaa aacactacac gatgctttaa
     6241 ctgcaaaaat tcagctcacc agttttgagg caaaattttt gagtgacatg caaagtaagt
     6301 atgatctcaa tggttcgttc tcatggctca cgcaaaaaca acgaaccaca ctagataaca
     6361 tactggctaa atacggaagg atctgaggtt cttatggctc ttgtatctat cagtgaagca
     6421 tcaagactaa caaacaaaag tagaacaact gttcaccgtt acatatcaaa gggaaaactg
     6481 tccatatgca cagatgaaaa cggtgtaaaa aagatagata catcagagct tttacgagtt
     6541 tttggtgcat tcaaagctgt tcaccatgaa cagatcgaca atgtaacaga tgaacagcat
     6601 gtaacaccta atagaacagg tgaaaccagt aaaacaaagc aactagaaca tgaaattgaa
     6661 cacctgagac aacttgttac agctcaacag tcacacatag acagcctgaa acaggcgatg
     6721 ctgcttatcg aatcaaagct gccgacaaca cgggagccag tgacgcctcc cgtggggaaa
     6781 aaatcatggc aattctggaa gaaatagcgc tttcagccgg caaaccggct gaagccggat
     6841 ctgcgattct gataacaaac tagcaacacc agaacagccc gtttgcgggc agcaaaaccc
     6901 gtacttttgg acgttccggc ggttttttgt ggcgagtggt gttcgggcgg tgcgcgcaag
     6961 atccattatg ttaaacgggc gagtttacat ctcaaaaccg cccgcttaac accatcagaa
     7021 atcctcagcg cgattttaag caccaacccc cccccgtaac acccaaatcc atactgaaag
     7081 tggctttgtt gaataaatca gatttcgggt aagtctcccc cgtagcgggt tgtgttttca
     7141 ggcaatacgc acgctttcag gcatacctgc tttcgtcatt ttgttcagcg ctcgtaccag
     7201 ggccatagcc tccgcaacct gaccatcgta gtcacgcagc gtcagtgaac ccccgaacag
     7261 agatctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     7321 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     7381 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     7441 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     7501 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     7561 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     7621 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     7681 ag
//
