LOCUS       sequence-388085-        7227 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Epitope tagging of effector protein PipB2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7227)
  TITLE     Hensel lab plasmids
REFERENCE   2  (bases 1 to 7227)
  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: p2621
FEATURES             Location/Qualifiers
     source          1..7227
                     /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"
     polyA_signal    4007..4128
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4018..4037)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4072..4091
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5829..5845)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5830..5846)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(5870..5889)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5871..5889)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5899..5916)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5899..5915)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5908..5930)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6057..6512
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6144..6163)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6354..6375
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6538..6642
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6643..6711,6712..7227,1..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(6861..6880)
                     /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 aaggcttaag 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 caaatccgac 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 agtcgaccag acatgataag atacattgat
     4021 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     4081 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttct gctttaataa
     4141 gatcctagaa ctagaggatc cgcgtctctg tctcaaaagg aggtaggcga tccctactcc
     4201 gatccatggc accaccccca tcaatatttt cactataaaa ttcgttaaag agtgtttgtg
     4261 tgcttgtaga cattgtggcg tcttcagtac gcaaatcggt tagtgtacaa tctttaaaac
     4321 aggtattgtt taaattagcc tcagccaaat cagccccgtc caggttcgta gaaatgaaac
     4381 tcgcctttcg caggtttgcc cgggttaaga cagcgccttc cataattgca gccattatag
     4441 tagcaccctg taatgtcgca ccgctcatat cagcacgtat aagggttgcg cctgaaagat
     4501 tagtgtgatc catatttgca ccggagcaat tacattctat catgctacag cctaacagac
     4561 tcgcgtgttg taaattggcg ccagtaagat tagcgtgatc gagggtagcg ccacacataa
     4621 tgctatcttc cagacaggcg ttcttgaaat tactgtcgcc tagtgatgtt cccgatagat
     4681 ttgcgccctt aaaatttgca ccttcacaat ccgccataaa cagcactgcc ccctctaaat
     4741 ttgcgccgct taagtctgcg tcacataaat tttctgacgt cagatttgct ccggtaagga
     4801 cagcgccttt aagattcatg cctccctggg cagttaatat tacagaatcc tgagggagag
     4861 aaaattcgca tctgaatagt aaagctcggc atattttttc aaaagaggca agcggaattt
     4921 ctcttgtctc agaatggtcg cctttactga ctcgtacaat aacttgtcca gcttcgttat
     4981 tctcaccagg aagattaaat tcgacacgac acccatccat atcatccagg atgatgtttt
     5041 ctggcaacgg agcacctctg tcaggcattg tacttttcaa tgtctcagcc atagtctcta
     5101 tcaattcctg atattgtgtt tcatttctcc gacgtatccc accacaggta aaaaagttaa
     5161 tgatatattc cagaatggtt ttgggcgagg tagcttgccg catagcagca gaagtccccg
     5221 cgccaaaagc agatttagcc ataccagcca gactatcgag tgaacgctcc atatattttc
     5281 tcccagagac agcaacacag tgcatgataa aatttatcat atagatttgc atttagagat
     5341 aaggtgaata ccccctttac gtctggttac ataataacgt atcacgacgc gttgctgtct
     5401 ggcttacaca atagcagcca aataaatata ttggcttata ctcgtcatac ttcaagttgc
     5461 atgtgctgcg gccgcgttcc ctcaccccag tcacttactt tagtaagctc ctggggattc
     5521 gctcgcttgc cgccttcctg caactcgaat tatttagagt atataacatc atttaaaata
     5581 ataaacacgg cattcgtttt aatactacgt tctggactca tcccactcat tagcagaaaa
     5641 cctattaaaa agcgtgttca ggcattttat taccgccatt gataaactgt ttaacaacat
     5701 cgtctgtaca gaccttcttc gttgccttta cgtttaactc aatcaggcta ccgtctcggt
     5761 tataagtcta tcgagtagta gagccgtagt atgacgatgc agcaaagtga attcgatatc
     5821 aagcttatcg ataccgtcga cctcgagggg gggcccggta cccaattcgc cctatagtga
     5881 gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     5941 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     6001 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc
     6061 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     6121 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     6181 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     6241 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     6301 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     6361 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     6421 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     6481 gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc
     6541 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     6601 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     6661 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     6721 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     6781 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     6841 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     6901 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     6961 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     7021 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     7081 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgctg ggaaccggag
     7141 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     7201 acgttgcgca aactattaac tggcgaa
//
