LOCUS       sequence-415171-        4682 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Template for hlpA-mkate2 amplification associated with erythromycin 
            resistance gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4682)
  AUTHORS   Mignolet J, Staub J, van der Meer JR, Veening J-W, Libante V
  TITLE     A Modular Genetic Toolbox for Precise Gene Regulation and 
            Multi-Color Imaging in Streptococci
  JOURNAL   bioRxiv 2025.05.10.653270
REFERENCE   2  (bases 1 to 4682)
  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"; journalName: "bioRxiv 
            2025.05.10.653270"
COMMENT     Sequence Label: pJethlpAmKate2Ery
FEATURES             Location/Qualifiers
     source          1..4682
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     305..324
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        305..323
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(join(2514..2520,2521..2538,2539..2544))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    2559..2580
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2700..2717)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2871..3459)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2951..2970)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3630..4421,4422..4490))
                     /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     4253..4272
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4491..4595)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4663..4681
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gcccctgcag ccgaattata ttatttttgc caaataattt ttaacaaaag ctctgaagtc
       61 ttcttcattt aaattcttag atgatacttc atctggaaaa ttgtcccaat tagtagcatc
      121 acgctgtgag taagttctaa accatttttt tattgttgta ttatctctaa tcttactact
      181 cgatgagttt tcggtattat ctctattttt aacttggagc aggttccatt cattgttttt
      241 ttcatcatag tgaataaaat caactgcttt aacacttgtg cctgaacacc atatccatcc
      301 ggcgtaatac gactcactat agggagagcg gccgccagat cttccggatg gctcgagttt
      361 ttcagcaaga tcatggcaaa caaacaagat ttgatcgcta aagtagcaga agctacagaa
      421 ttgactaaga aagactcagc agcagcagtt gaagctgtat ttgcagcagt agctgactat
      481 cttgcagctg gtgaaaaagt tcaattgatc ggttttggta actttgaagt tcgtgagcgt
      541 gcagaacgta aaggtcgcaa cccacaaact ggtaaagaaa tgacaattgc agcttctaaa
      601 gtaccagcat tcaaagctgg taaagctctt aaagacgctg ttaaaggatc cggatctggt
      661 ggagaagctg cagctaaagg atcagaactt atcaaggaaa atatgcacat gaaattgtac
      721 atggaaggaa cagtaaataa tcaccacttt aaatgtacct cagaaggaga aggaaaacca
      781 tatgaaggta ctcaaaccat gcgtattaag gccgttgaag gtggaccatt gccttttgcc
      841 tttgatattc ttgctacatc ttttatgtac ggatcaaaga cttttatcaa tcatacccaa
      901 ggtatcccag atttctttaa acagtcattt cctgaaggat ttacatggga acgtgtcaca
      961 acttatgaag atggtggagt attgacagca actcaagata catctttgca agatggttgt
     1021 cttatctaca atgtaaagat ccgtggagtt aattttccaa gtaatggtcc tgttatgcag
     1081 aaaaagaccc ttggatggga agcatctacc gaaacattat atcctgccga tggtggattg
     1141 gaaggtcgtg ctgatatggc attgaaactt gtcggtggag gtcaccttat ctgtaatttg
     1201 aagaccacat accgttctaa aaagccagct aagaatctta agatgcctgg tgtttactac
     1261 gtcgatcgtc gtttagaacg tatcaaagaa gcagataagg aaacttatgt tgaacagcac
     1321 gaagtagccg tcgcacgtta ttgtgatttg cctagtaaat tgggacaccg ttaataagaa
     1381 ttccaaactt aagagtgtgt tgatagtgca gtatcttaaa attttgtata ataggaattg
     1441 aagttaaatt agatgctaaa aatttgtaat taagaaggag tgattacatg aacaaaaata
     1501 taaaatattc tcaaaacttt ttaacgagtg aaaaagtact caaccaaata ataaaacaat
     1561 tgaatttaaa agaaaccgat accgtttacg aaattggaac aggtaaaggg catttaacga
     1621 cgaaactggc taaaataagt aaacaggtaa cgtctattga attagacagt catctattca
     1681 acttatcgtc agaaaaatta aaactgaata ctcgtgtcac tttaattcac caagatattc
     1741 tacagtttca attccctaac aaacagaggt ataaaattgt tgggagtatt ccttaccatt
     1801 taagcacaca aattattaaa aaagtggttt ttgaaagcca tgcgtctgac atctatctga
     1861 ttgttgaaga aggattctac aagcgtacct tggatattca ccgaacacta gggttgctct
     1921 tgcacactca agtctcgatt cagcaattgc ttaagctgcc agcggaatgc tttcatccta
     1981 aaccaaaagt aaacagtgtc ttaataaaac ttacccgcca taccacagat gttccagata
     2041 aatattggaa gctatatacg tactttgttt caaaatgggt caatcgagaa tatcgtcaac
     2101 tgtttactaa aaatcagttt catcaagcaa tgaaacacgc caaagtaaac aatttaagta
     2161 ccgttactta tgagcaagta ttgtctattt ttaatagtta tctattattt aacgggagga
     2221 aataattcta tgagtcgctt ttgtaaattt ggaaagttac acgttactaa agggaatgta
     2281 gataaattat taggtatact actgacagct tccaaggagc taaagaggtc caagcttcag
     2341 gaaactgaga caggaatttt attaaaaatt taaattttga agaaagttca gggttaatag
     2401 catccatttt ttgctttgca agttcctcag cattcttaac aaaagacgtc tcttttgaca
     2461 tgtttaaagt ttaaacctcc tgtgtgaaat tattatccgc tcataattcc acacattata
     2521 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     2581 attgcgttgc gctcactgcc aattgctttc cagtcgggaa acctgtcgtg ccagctgcat
     2641 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     2701 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     2761 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     2821 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     2881 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     2941 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     3001 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     3061 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     3121 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     3181 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     3241 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     3301 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     3361 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     3421 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     3481 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     3541 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     3601 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     3661 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     3721 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     3781 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     3841 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     3901 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     3961 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     4021 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     4081 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     4141 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     4201 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     4261 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     4321 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     4381 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     4441 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     4501 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     4561 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     4621 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     4681 cc
//
