LOCUS       sequence-415030-        4918 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Template for hlpA-mkate2 amplification associated with 
            chloramphenicol resistance gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4918)
  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 4918)
  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: pJethlpAmKate2Cm
FEATURES             Location/Qualifiers
     source          1..4918
                     /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(2750..2756,2757..2774,2775..2780))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    2795..2816
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2936..2953)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3107..3695)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3187..3206)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3866..4657,4658..4726))
                     /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     4489..4508
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4727..4831)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4899..4917
                     /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 ttccgtattg gtggaataac acgaattaat aaaggatata tttgtagtag caagtgtatt
     1441 tgttttatat tctatgaccc tatttgattt acatgattct atatggtata atggacttgc
     1501 ttataaatat agatgcttgg atagcaggag gttgcaagat gatgatggca gttaaattag
     1561 ccttctaaaa ttaaataata aaattttagg aggctaattt atgaaattta atttgataga
     1621 tatagaggat tggaatagaa agccatactt tgagcattat ttaaatgcgg taaggtgcac
     1681 ttacagtatg actgcaaatt tagagataac tagtttactg cgtgaaatta aacttaaggg
     1741 cctgaaactg taccctacgc ttatttatat catcacaact gtagttaatc gtcacaagga
     1801 gttccgcacc agttttgatc aaaacggtaa gttaggatac tgggatagta tgaacccaag
     1861 ttatactatc tttcataagg ataacgaaac tttttcaagt atttggacag agtatgacga
     1921 gaacttccca cgtttctact ataattacct tgaggatatc agaaactata gcgacgttat
     1981 gaatttcatg cctaaggcag gtcaacctgc taacacaatt aatgtgtcca gcattccttg
     2041 ggtgaatttt accggattca acttgaatat atataatgat gcaacatatc taatccctat
     2101 ttttactatg ggtaagtatt ttcagcagga taataaaatt ttattaccta tgtctgtaca
     2161 ggtgcatcat gcggtttgcg acggttacca taccagcaga ttttttaatg aggtacagga
     2221 attagtctca aattttgaga catggttagg agaaaaataa tattaaaatg cttgtgctcg
     2281 ggagttataa acaaagatcg aaaatattgt taagatcgtg aggaagcaaa gcgaccgaag
     2341 aagtatatgt tccatctatg caaggacttc ccgagaaggc gagtgacagc gaagaccgat
     2401 tggaaggagt ctgagctaag tggagtaata tattcattca ttatagaacc atcatttcaa
     2461 aggattagag aaatctaatc ctttttctag gtatttttat cctaagaatt taatatgaca
     2521 atcacgagtt taatttttat ttgggtgcta tttatcctaa gcagagcaag cttcaggaaa
     2581 ctgagacagg aattttatta aaaatttaaa ttttgaagaa agttcagggt taatagcatc
     2641 cattttttgc tttgcaagtt cctcagcatt cttaacaaaa gacgtctctt ttgacatgtt
     2701 taaagtttaa acctcctgtg tgaaattatt atccgctcat aattccacac attatacgag
     2761 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     2821 cgttgcgctc actgccaatt gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     2881 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     2941 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     3001 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     3061 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     3121 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     3181 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     3241 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     3301 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     3361 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     3421 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     3481 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     3541 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     3601 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     3661 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     3721 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     3781 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     3841 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     3901 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     3961 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     4021 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     4081 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     4141 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     4201 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     4261 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     4321 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     4381 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     4441 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     4501 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     4561 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     4621 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     4681 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     4741 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     4801 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     4861 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggcc
//
