LOCUS       sequence-415203-        4688 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Template for hlpA-mScarlet-I amplification associated with 
            erythromycin resistance gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4688)
  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 4688)
  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: pJethlpAmScarletEry
FEATURES             Location/Qualifiers
     source          1..4688
                     /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(2520..2526,2527..2544,2545..2550))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    2565..2586
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2706..2723)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2877..3465)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2957..2976)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3636..4427,4428..4496))
                     /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     4259..4278
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4497..4601)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4669..4687
                     /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 atcagtgagt aaaggcgaag ctgtaataaa ggagttcatg
      721 agatttaaag tacacatgga ggggtcaatg aacggtcatg aatttgaaat tgagggagag
      781 ggagagggaa gaccgtatga gggtacgcaa accgccaaat tgaaggtaac gaagggggga
      841 cctcttcctt tttcatggga cattcttagc ccgcagttca tgtatgggag ccgagcattt
      901 attaagcacc cagcggacat accagattat tacaaacaat ctttccctga aggtttcaag
      961 tgggagagag ttatgaactt tgaggacgga ggagctgtaa cagtgaccca agatacttca
     1021 ctagaggacg ggactcttat ttacaaagtc aaactacgag gaacgaactt tccgcctgac
     1081 ggccctgtta tgcagaaaaa gactatgggc tgggaagcct ctacggagcg cctataccca
     1141 gaggatggtg ttttgaaagg tgacattaaa atggcacttc gcctaaagga cggaggccga
     1201 tacctagcag attttaaaac aacgtataaa gctaagaaac ctgtacaaat gccaggagca
     1261 tataatgttg atcgaaaatt agatattact tctcataacg aggactacac agtggttgaa
     1321 caatatgaac gtagcgaagg tcgtcattca actgggggca tggacgaact atataaataa
     1381 taagaattcc aaacttaaga gtgtgttgat agtgcagtat cttaaaattt tgtataatag
     1441 gaattgaagt taaattagat gctaaaaatt tgtaattaag aaggagtgat tacatgaaca
     1501 aaaatataaa atattctcaa aactttttaa cgagtgaaaa agtactcaac caaataataa
     1561 aacaattgaa tttaaaagaa accgataccg tttacgaaat tggaacaggt aaagggcatt
     1621 taacgacgaa actggctaaa ataagtaaac aggtaacgtc tattgaatta gacagtcatc
     1681 tattcaactt atcgtcagaa aaattaaaac tgaatactcg tgtcacttta attcaccaag
     1741 atattctaca gtttcaattc cctaacaaac agaggtataa aattgttggg agtattcctt
     1801 accatttaag cacacaaatt attaaaaaag tggtttttga aagccatgcg tctgacatct
     1861 atctgattgt tgaagaagga ttctacaagc gtaccttgga tattcaccga acactagggt
     1921 tgctcttgca cactcaagtc tcgattcagc aattgcttaa gctgccagcg gaatgctttc
     1981 atcctaaacc aaaagtaaac agtgtcttaa taaaacttac ccgccatacc acagatgttc
     2041 cagataaata ttggaagcta tatacgtact ttgtttcaaa atgggtcaat cgagaatatc
     2101 gtcaactgtt tactaaaaat cagtttcatc aagcaatgaa acacgccaaa gtaaacaatt
     2161 taagtaccgt tacttatgag caagtattgt ctatttttaa tagttatcta ttatttaacg
     2221 ggaggaaata attctatgag tcgcttttgt aaatttggaa agttacacgt tactaaaggg
     2281 aatgtagata aattattagg tatactactg acagcttcca aggagctaaa gaggtccaag
     2341 cttcaggaaa ctgagacagg aattttatta aaaatttaaa ttttgaagaa agttcagggt
     2401 taatagcatc cattttttgc tttgcaagtt cctcagcatt cttaacaaaa gacgtctctt
     2461 ttgacatgtt taaagtttaa acctcctgtg tgaaattatt atccgctcat aattccacac
     2521 attatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     2581 acattaattg cgttgcgctc actgccaatt gctttccagt cgggaaacct gtcgtgccag
     2641 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     2701 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     2761 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     2821 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     2881 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     2941 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     3001 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     3061 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     3121 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     3181 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     3241 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     3301 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     3361 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     3421 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     3481 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     3541 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     3601 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     3661 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     3721 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     3781 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     3841 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     3901 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     3961 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     4021 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     4081 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     4141 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     4201 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     4261 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     4321 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     4381 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     4441 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     4501 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     4561 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     4621 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     4681 acgaggcc
//
