LOCUS       sequence-415308-        4822 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Template for hlpA-mScarlet-I amplification associated with 
            spectinomycin resistance gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4822)
  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 4822)
  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: pJethlpAmScarletSpc
FEATURES             Location/Qualifiers
     source          1..4822
                     /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(2654..2660,2661..2678,2679..2684))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    2699..2720
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2840..2857)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3011..3599)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3091..3110)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3770..4561,4562..4630))
                     /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     4393..4412
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4631..4735)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4803..4821
                     /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 gaagttatcg taacgtgact ggcaagagat atttttaaaa caatgaatag
     1441 gtttacactt actttagttt tatggaaatg aaagatcata tcatatataa tctagaataa
     1501 aattaactaa aataattatt atctagataa aaaatttaga agccaatgaa atctataaat
     1561 aaactaaatt aagtttattt aattaacaac tatggatata aaataggtac taatcaaaat
     1621 agtgaggagg atatatttga atacatacga acaaattaat aaagtgaaaa aaatacttcg
     1681 gaaacattta aaaaataacc ttattggtac ttacatgttt ggatcaggag ttgagagtgg
     1741 actaaaacca aatagtgatc ttgacttttt agtcgtcgta tctgaaccat tgacagatca
     1801 aagtaaagaa atacttatac aaaaaattag acctatttca aaaaaaatag gagataaaag
     1861 caacttacga tatattgaat taacaattat tattcagcaa gaaatggtac cgtggaatca
     1921 tcctcccaaa caagaattta tttatggaga atggttacaa gagctttatg aacaaggata
     1981 cattcctcag aaggaattaa attcagattt aaccataatg ctttaccaag caaaacgaaa
     2041 aaataaaaga atatacggaa attatgactt agaggaatta ctacctgata ttccattttc
     2101 tgatgtgaga agagccatta tggattcgtc agaggaatta atagataatt atcaggatga
     2161 tgaaaccaac tctatattaa ctttatgccg tatgatttta actatggaca cgggtaaaat
     2221 cataccaaaa gatattgcgg gaaatgcagt ggctgaatct tctccattag aacataggga
     2281 gagaattttg ttagcagttc gtagttatct tggagagaat attgaatgga ctaatgaaaa
     2341 tgtaaattta actataaact atttaaataa cagattaaaa aaattataaa aaaattgaaa
     2401 aaatggtgga aacacttttt taattttttt gttttattat ttaatatttg ggaaatattc
     2461 attctaattg gaagcttcag gaaactgaga caggaatttt attaaaaatt taaattttga
     2521 agaaagttca gggttaatag catccatttt ttgctttgca agttcctcag cattcttaac
     2581 aaaagacgtc tcttttgaca tgtttaaagt ttaaacctcc tgtgtgaaat tattatccgc
     2641 tcataattcc acacattata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     2701 gagtgagcta actcacatta attgcgttgc gctcactgcc aattgctttc cagtcgggaa
     2761 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     2821 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     2881 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     2941 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     3001 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     3061 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     3121 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     3181 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     3241 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     3301 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     3361 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     3421 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     3481 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     3541 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     3601 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     3661 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     3721 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     3781 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     3841 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     3901 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     3961 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     4021 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     4081 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     4141 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     4201 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     4261 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     4321 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     4381 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     4441 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     4501 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     4561 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     4621 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     4681 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     4741 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     4801 aaaataggcg tatcacgagg cc
//
