LOCUS       sequence-388082-        7133 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bacterial expression of sseF::3xHA fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7133)
  TITLE     Hensel lab plasmids
REFERENCE   2  (bases 1 to 7133)
  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: p5065
FEATURES             Location/Qualifiers
     source          1..7133
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1298..2248)
                     /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      2296..2518
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     primer_bind     3663..3680
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3797..3818
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3833..3838,3839..3856,3857..3863)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3871..3887
                     /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     3876..3898
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3895..3911
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3895..3911
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3930..3950
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3932..3950
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             complement(3996..4085)
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     primer_bind     complement(5735..5751)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5736..5752)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(5776..5795)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5777..5795)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5805..5822)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5805..5821)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5814..5836)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5963..6418
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6050..6069)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6260..6281
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6444..6548
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6549..6617,6618..7133,1..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(6767..6786)
                     /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 gggggggttg gtgcttaaaa tcgcgctgag
      661 gatttctgat ggtgttaagc gggcggtttt gagatgtaaa ctcgcccgtt taacataatg
      721 gatcttgcgc gcaccgcccg aacaccactc gccacaaaaa accgccggaa cgtccaaaag
      781 tacgggtttt gctgcccgca aacgggctgt tctggtgttg ctagtttgtt atcagaatcg
      841 cagatccggc ttcagccggt ttgccggctg aaagcgctat ttcttccaga attgccatga
      901 ttttttcccc acgggaggcg tcactggctc ccgtgttgtc ggcagctttg attcgataag
      961 cagcatcgcc tgtttcaggc tgtctatgtg tgactgttga gctgtaacaa gttgtctcag
     1021 gtgttcaatt tcatgttcta gttgctttgt tttactggtt tcacctgttc tattaggtgt
     1081 tacatgctgt tcatctgtta cattgtcgat ctgttcatgg tgaacagctt tgaatgcacc
     1141 aaaaactcgt aaaagctctg atgtatctat cttttttaca ccgttttcat ctgtgcatat
     1201 ggacagtttt ccctttgata tgtaacggtg aacagttgtt ctacttttgt ttgttagtct
     1261 tgatgcttca ctgatagata caagagccat aagaacctca gatccttccg tatttagcca
     1321 gtatgttctc tagtgtggtt cgttgttttt gcgtgagcca tgagaacgaa ccattgagat
     1381 catacttact ttgcatgtca ctcaaaaatt ttgcctcaaa actggtgagc tgaatttttg
     1441 cagttaaagc atcgtgtagt gtttttctta gtccgttatg taggtaggaa tctgatgtaa
     1501 tggttgttgg tattttgtca ccattcattt ttatctggtt gttctcaagt tcggttacga
     1561 gatccatttg tctatctagt tcaacttgga aaatcaacgt atcagtcggg cggcctcgct
     1621 tatcaaccac caatttcata ttgctgtaag tgtttaaatc tttacttatt ggtttcaaaa
     1681 cccattggtt aagcctttta aactcatggt agttattttc aagcattaac atgaacttaa
     1741 attcatcaag gctaatctct atatttgcct tgtgagtttt cttttgtgtt agttctttta
     1801 ataaccactc ataaatcctc atagagtatt tgttttcaaa agacttaaca tgttccagat
     1861 tatattttat gaattttttt aactggaaaa gataaggcaa tatctcttca ctaaaaacta
     1921 attctaattt ttcgcttgag aacttggcat agtttgtcca ctggaaaatc tcaaagcctt
     1981 taaccaaagg attcctgatt tccacagttc tcgtcatcag ctctctggtt gctttagcta
     2041 atacaccata agcattttcc ctactgatgt tcatcatctg agcgtattgg ttataagtga
     2101 acgataccgt ccgttctttc cttgtagggt tttcaatcgt ggggttgagt agtgccacac
     2161 agcataaaat tagcttggtt tcatgctccg ttaagtcata gcgactaatc gctagttcat
     2221 ttgctttgaa aacaactaat tcagacatac atctcaattg gtctaggtga ttttaatcac
     2281 tataccaatt gagatgggct agtcaatgat aattactagt ccttttcctt tgagttgtgg
     2341 gtatctgtaa attctgctag acctttgctg gaaaacttgt aaattctgct agaccctctg
     2401 taaattccgc tagacctttg tgtgtttttt ttgtttatat tcaagtggtt ataatttata
     2461 gaataaagaa agaataaaaa aagataaaaa gaatagatcc cagccctgtg tataactcac
     2521 tactttagtc agttccgcag tattacaaaa ggatgtcgca aacgctgttt gctcctctac
     2581 aaaacagacc ttaaaaccct aaaggcttaa gtagcaccct cgcaagctcg ggcaaatcgc
     2641 tgaatattcc ttttgtctcc gaccatcagg cacctgagtc gctgtctttt tcgtgacatt
     2701 cagttcgctg cgctcacggc tctggcagtg aatgggggta aatggcacta caggcgcctt
     2761 ttatggattc atgcaaggaa actacccata atacaagaaa agcccgtcac gggcttctca
     2821 gggcgtttta tggcgggtct gcggagtggt gaatccgtta gcgaggtgcc gccggcttcc
     2881 attcaggtcg aggtggcccg gctccatgca ccgcgacgca acgcggggag gcagacaagg
     2941 tatagggcgg cgcctacaat ccatgccaac ccgttccatg tgctcgccga ggcggcataa
     3001 atcgccgtga cgatcagcgg tccaatgatc gaagttaggc tggtaagagc cgcgagcgat
     3061 ccttgaagct gtccctgatg gtcgtcatct acctgcctgg acagcatggc ctgcaacgcg
     3121 ggcatcccga tgccgccgga agcgagaaga atcataatgg ggaaggccat ccagcctcgc
     3181 gtcgcgaacg ccagcaagac gtagcccagc gcgtcggccg ccatgccggc gataatggcc
     3241 tgcttctcgc cgaaacgttt ggtggcggga ccagtgacga aggcttgagc gagggcgtgc
     3301 aagattccga ataccgcaag cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc
     3361 tcgccgaaaa tgacccagag cgctgccggc accctgtcct acgagttgca tgataaagaa
     3421 gacagtcata agtgcggcga cgatagtcat gccccgcgcc caccggaagg agctgactgg
     3481 gttgaaggct ctcaagggca tcggcattcg ccattcaggc tgcctcctac caagacgact
     3541 tcaaatccga cccgtagaga tctcacatgt tctttcctgc gttatcccct gattctgtgg
     3601 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     3661 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     3721 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     3781 gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact
     3841 ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
     3901 acagctatga ccatgattac gccaagcgcg caattaaccc tcactaaagg gaacaaaagc
     3961 tggagctcca ccgcggtggc ggccgctcta gattaagcgt aatctggaac gtcatatgga
     4021 taggatcctg catagtccgg gacgtcatag ggatagccag cgtagtctgg gacgtcgtat
     4081 gggtatggtt ctccccgaga tgtatgatca gaactgtcca ttaatgcagg tgtagtagca
     4141 gattgacaga gcgctgccat ttgttccgcc aataacacac tgggatcggc atgaagttca
     4201 tcaacagaat tttcctgatc gtcgccagag gggcgggcaa ggcaataatc cagtaccgca
     4261 cctatcgccg tcaggctaac ggaggtaatt acactcccca tgtccaaaga ggccgcaata
     4321 ttttcagccg cgggcagtgg aaactgtagg ggtaaaacca acatagaaat agcgattcct
     4381 gaacgtatta ataaagaaag acaattagca agggtgttag cgcagttaag acttgcccca
     4441 cattttaagg ccagcgcact gaccacaaga gcaacgctat cactggcggt ttgtaatggc
     4501 tccttttgct gacatatcga ttgataatta tgatacgcac agcaagcatc cccaatagca
     4561 atcacgagcg ccgcccccgc aagaatagca atgggtaatc ctgccccgcc agaaattacc
     4621 gctgcagcaa ccgataaccc aaacacgact gtcgcaccca gcgcacgaat agtgtattgc
     4681 ataaaatgta tcgcataatc cctctgctgc cttatttgtt caggcgtaag cagcacaggg
     4741 gctgcggggg tgccaggcgc tggaatttca gggggaggaa acgatacctc cttcgcttgt
     4801 atatcggaag gaggactatt accatcgact atattacttg ccgctgacgg aatatgaatt
     4861 ttcatatttc gttctgttat ttaagcaata agagtatcaa ccattatttg cgcattctgg
     4921 cgaatctcac tccatgaggc atccgcataa ctcatcttga ttgcggtttg aaaagcctct
     4981 ctcgccaacc cgggttcccc catcattttg agacagacgc ccgtttggta aaccggttct
     5041 ggatggctgg catccagcat caaggcatgt ccatagaaat taatggccgt cgtgtattct
     5101 ttaagcatca tccaggtgcc agccaatgca atatgggcac gccaactcca tggctgggcc
     5161 atcaccagcc aactaaaatc gattacggcg cgcgaataat ccccctcctg ccatgaggcg
     5221 taaccactgg cataaacggt ttccggatca acggataata gctgtttcag aatgtcttcg
     5281 ggtattttat ttttctgatc ttctttcatc atcataccta ttgaattcac gataagataa
     5341 ttaacgtgct aactaagagc ctatcccatt agggctattt tacttgccat tttgaacctg
     5401 ggcagtgctc gccaaaacgc gttagcgttt tgaacgccgc tagcggcggc ccgaagggcg
     5461 agcgtagcga gtcaaacctc acgtactacg tgtacgctgc ggtttttgcg cgctatccgt
     5521 gtccaaactg gctgcgccaa ttaacgcctg gtgggatagg ctctaagata tttttacttt
     5581 actcttgctc actcactaca agtgcgctgt tatggtaacg ataataaata atgttgatga
     5641 tattttcggc ctgctcgatc gcttcaagca ataaggtgtt ttgctgatat tgctgcggat
     5701 cctgtaactt gccgttgttc tcttctaagc ttatcgatac cgtcgacctc gagggggggc
     5761 ccggtaccca attcgcccta tagtgagtcg tattacgcgc gctcactggc cgtcgtttta
     5821 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     5881 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     5941 cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     6001 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     6061 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     6121 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     6181 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     6241 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     6301 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     6361 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag
     6421 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     6481 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     6541 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     6601 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     6661 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     6721 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     6781 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     6841 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     6901 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     6961 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     7021 ctcgccttga tcgctgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     7081 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaa
//
