LOCUS       sequence-415199-        4924 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Template for hlpA-mScarlet-I amplification associated with 
            chloramphenicol resistance gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4924)
  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 4924)
  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: pJethlpAmScarletCm
FEATURES             Location/Qualifiers
     source          1..4924
                     /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(2756..2762,2763..2780,2781..2786))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    2801..2822
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2942..2959)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3113..3701)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3193..3212)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3872..4663,4664..4732))
                     /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     4495..4514
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4733..4837)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4905..4923
                     /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 gtattggtgg aataacacga attaataaag gatatatttg tagtagcaag
     1441 tgtatttgtt ttatattcta tgaccctatt tgatttacat gattctatat ggtataatgg
     1501 acttgcttat aaatatagat gcttggatag caggaggttg caagatgatg atggcagtta
     1561 aattagcctt ctaaaattaa ataataaaat tttaggaggc taatttatga aatttaattt
     1621 gatagatata gaggattgga atagaaagcc atactttgag cattatttaa atgcggtaag
     1681 gtgcacttac agtatgactg caaatttaga gataactagt ttactgcgtg aaattaaact
     1741 taagggcctg aaactgtacc ctacgcttat ttatatcatc acaactgtag ttaatcgtca
     1801 caaggagttc cgcaccagtt ttgatcaaaa cggtaagtta ggatactggg atagtatgaa
     1861 cccaagttat actatctttc ataaggataa cgaaactttt tcaagtattt ggacagagta
     1921 tgacgagaac ttcccacgtt tctactataa ttaccttgag gatatcagaa actatagcga
     1981 cgttatgaat ttcatgccta aggcaggtca acctgctaac acaattaatg tgtccagcat
     2041 tccttgggtg aattttaccg gattcaactt gaatatatat aatgatgcaa catatctaat
     2101 ccctattttt actatgggta agtattttca gcaggataat aaaattttat tacctatgtc
     2161 tgtacaggtg catcatgcgg tttgcgacgg ttaccatacc agcagatttt ttaatgaggt
     2221 acaggaatta gtctcaaatt ttgagacatg gttaggagaa aaataatatt aaaatgcttg
     2281 tgctcgggag ttataaacaa agatcgaaaa tattgttaag atcgtgagga agcaaagcga
     2341 ccgaagaagt atatgttcca tctatgcaag gacttcccga gaaggcgagt gacagcgaag
     2401 accgattgga aggagtctga gctaagtgga gtaatatatt cattcattat agaaccatca
     2461 tttcaaagga ttagagaaat ctaatccttt ttctaggtat ttttatccta agaatttaat
     2521 atgacaatca cgagtttaat ttttattttg gtgctattta tcctaagcag agcaagcttc
     2581 aggaaactga gacaggaatt ttattaaaaa tttaaatttt gaagaaagtt cagggttaat
     2641 agcatccatt ttttgctttg caagttcctc agcattctta acaaaagacg tctcttttga
     2701 catgtttaaa gtttaaacct cctgtgtgaa attattatcc gctcataatt ccacacatta
     2761 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     2821 taattgcgtt gcgctcactg ccaattgctt tccagtcggg aaacctgtcg tgccagctgc
     2881 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     2941 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     3001 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     3061 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     3121 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     3181 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     3241 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     3301 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     3361 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     3421 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     3481 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     3541 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     3601 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     3661 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     3721 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     3781 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     3841 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     3901 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     3961 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     4021 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     4081 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     4141 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     4201 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     4261 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     4321 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     4381 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     4441 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     4501 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     4561 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     4621 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     4681 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     4741 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     4801 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     4861 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     4921 ggcc
//
