LOCUS       sequence-415321-        4977 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Template for hlpA-YFP amplification associated with chloramphenicol 
            resistance gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4977)
  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 4977)
  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: pJethlpAYFPCm
FEATURES             Location/Qualifiers
     source          1..4977
                     /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(2809..2815,2816..2833,2834..2839))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    2854..2875
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2995..3012)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3166..3754)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3246..3265)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3925..4716,4717..4785))
                     /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     4548..4567
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4786..4890)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4958..4976
                     /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 cagctaaatc tagttctaga ggatctggtg gagaagctgc agctaaagct
      721 ggatcaaaag gagaagaact ttttacaggt gtagtaccta tcttggttga attggatggt
      781 gatgttaacg gtcacaaatt ttctgtacgt ggtgaaggtg aaggtgatgc aactaacggt
      841 aaattgacac ttaaattgat ttgtacaact ggaaaacttc ctgttccttg gcctactctt
      901 gttacaacat tgggttatgg acttatgtgt tttgctcgtt atcctgatca tatgaaacgt
      961 cacgattttt ttaaatctgc tatgccagaa ggttatgtac aagaacgtac aatttcattt
     1021 aaagatgacg gaacatataa aacacgtgct gaagtaaaat tcgaaggtga cactcttgtt
     1081 aatcgtatcg aattgaaagg aatcgatttc aaagaagatg gtaacatttt gggacacaaa
     1141 cttgaataca acttcaactc tcataatgtt tatatcacag ctgacaaaca aaaaaacggt
     1201 attaaagcta attttaaaat tcgtcacaat gttgaagatg gaggagttca attggctgat
     1261 cattatcaac aaaatacacc aatcggagac ggaccagtat tgcttccaga taaccactac
     1321 ctttcttatc aatcaaaact ttcaaaagat cctaacgaaa aacgtgacca tatggtactt
     1381 cttgaatttg ttacagcagc aggtatcact cacggtatgg acgaacttta taaataagaa
     1441 ttccgtattg gtggataaca cgaattaata aaggatatat ttgtagtagc aagtgtattt
     1501 gttttatatt ctatgaccct atttgattta catgattcta tatggtataa tggacttgct
     1561 tataaatata gatgcttgga tagcaggagg ttgcaagatg atgatggcag ttaaattagc
     1621 cttctaaaat taaataataa aattttagga ggctaattta tgaaatttaa tttgatagat
     1681 atagaggatt ggaatagaaa gccatacttt gagcattatt taaatgcggt aaggtgcact
     1741 tacagtatga ctgcaaattt agagataact agtttactgc gtgaaattaa acttaagggc
     1801 ctgaaactgt accctacgct tatttatatc atcacaactg tagttaatcg tcacaaggag
     1861 ttccgcacca gttttgatca aaacggtaag ttaggatact gggatagtat gaacccaagt
     1921 tatactatct ttcataagga taacgaaact ttttcaagta tttggacaga gtatgacgag
     1981 aacttcccac gtttctacta taattacctt gaggatatca gaaactatag cgacgttatg
     2041 aatttcatgc ctaaggcagg tcaacctgct aacacaatta atgtgtccag cattccttgg
     2101 gtgaatttta ccggattcaa cttgaatata tataatgatg caacatatct aatccctatt
     2161 tttactatgg gtaagtattt tcagcaggat aataaaattt tattacctat gtctgtacag
     2221 gtgcatcatg cggtttgcga cggttaccat accagcagat tttttaatga ggtacaggaa
     2281 ttagtctcaa attttgagac atggttagga gaaaaataat attaaaatgc ttgtgctcgg
     2341 gagttataaa caaagatcga aaatattgtt aagatcgtga ggaagcaaag cgaccgaaga
     2401 agtatatgtt ccatctatgc aaggacttcc cgagaaggcg agtgacagcg aagaccgatt
     2461 ggaaggagtc tgagctaagt ggagtaatat attcattcat tatagaacca tcatttcaaa
     2521 ggattagaga aatctaatcc tttttctagg tatttttatc ctaagaattt aatatgacaa
     2581 tcacgagttt aatttttatt ttggtgctat ttatcctaag cagagcaagc ttcaggaaac
     2641 tgagacagga attttattaa aaatttaaat tttgaagaaa gttcagggtt aatagcatcc
     2701 attttttgct ttgcaagttc ctcagcattc ttaacaaaag acgtctcttt tgacatgttt
     2761 aaagtttaaa cctcctgtgt gaaattatta tccgctcata attccacaca ttatacgagc
     2821 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     2881 gttgcgctca ctgccaattg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     2941 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     3001 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     3061 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     3121 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     3181 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     3241 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     3301 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     3361 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     3421 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3481 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     3541 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     3601 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     3661 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     3721 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     3781 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     3841 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     3901 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     3961 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     4021 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     4081 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     4141 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     4201 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     4261 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     4321 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     4381 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     4441 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     4501 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     4561 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     4621 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     4681 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     4741 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     4801 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     4861 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     4921 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggcc
//
