LOCUS       sequence-415195-        4694 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Template for hlpA-mneongreen amplification associated with 
            erythromycin resistance gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4694)
  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 4694)
  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: pJethlpAmNeonGreenEry
FEATURES             Location/Qualifiers
     source          1..4694
                     /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(2526..2532,2533..2550,2551..2556))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    2571..2592
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2712..2729)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2883..3471)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2963..2982)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3642..4433,4434..4502))
                     /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     4265..4284
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4503..4607)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4675..4693
                     /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 atcagtctct gaagaagata atatggctag tttgcctgct
      721 acccacgaat tacacatttt tggatcaatc aatggagtcg attttgatat ggttggtcaa
      781 ggaaccggta atccaaatga tggatacgaa gaattgaatc ttaagagtac taagggtgat
      841 ttacagtttt ctccatggat tttggtccct catatcggat acggttttca ccaatacttg
      901 ccatatcctg atggaatgtc tccttttcag gctgcaatgg ttgatggatc aggttatcaa
      961 gtccatcgta caatgcagtt tgaagatggt gctagtctta ccgttaatta tcgttacaca
     1021 tatgaaggat ctcacattaa aggtgaagcc caagtaaagg gaacaggttt tccagcagat
     1081 ggacctgtta tgactaattc acttaccgcc gctgattggt gtcgtagtaa aaagacttat
     1141 ccaaatgata agacaatcat ctctactttt aagtggtcat acacaactgg aaatggtaaa
     1201 cgttatcgtt caactgctcg taccacatac acctttgcaa aaccaatggc agccaattac
     1261 cttaagaatc aacctatgta cgtctttcgt aagacagaac ttaagcattc taagactgaa
     1321 cttaatttta aggaatggca gaaggctttt actgatgtaa tgggtatgga tgaattgtat
     1381 aaataataag aattccaaac ttaagagtgt gttgatagtg cagtatctta aaattttgta
     1441 taataggaat tgaagttaaa ttagatgcta aaaatttgta attaagaagg agtgattaca
     1501 tgaacaaaaa tataaaatat tctcaaaact ttttaacgag tgaaaaagta ctcaaccaaa
     1561 taataaaaca attgaattta aaagaaaccg ataccgttta cgaaattgga acaggtaaag
     1621 ggcatttaac gacgaaactg gctaaaataa gtaaacaggt aacgtctatt gaattagaca
     1681 gtcatctatt caacttatcg tcagaaaaat taaaactgaa tactcgtgtc actttaattc
     1741 accaagatat tctacagttt caattcccta acaaacagag gtataaaatt gttgggagta
     1801 ttccttacca tttaagcaca caaattatta aaaaagtggt ttttgaaagc catgcgtctg
     1861 acatctatct gattgttgaa gaaggattct acaagcgtac cttggatatt caccgaacac
     1921 tagggttgct cttgcacact caagtctcga ttcagcaatt gcttaagctg ccagcggaat
     1981 gctttcatcc taaaccaaaa gtaaacagtg tcttaataaa acttacccgc cataccacag
     2041 atgttccaga taaatattgg aagctatata cgtactttgt ttcaaaatgg gtcaatcgag
     2101 aatatcgtca actgtttact aaaaatcagt ttcatcaagc aatgaaacac gccaaagtaa
     2161 acaatttaag taccgttact tatgagcaag tattgtctat ttttaatagt tatctattat
     2221 ttaacgggag gaaataattc tatgagtcgc ttttgtaaat ttggaaagtt acacgttact
     2281 aaagggaatg tagataaatt attaggtata ctactgacag cttccaagga gctaaagagg
     2341 tccaagcttc aggaaactga gacaggaatt ttattaaaaa tttaaatttt gaagaaagtt
     2401 cagggttaat agcatccatt ttttgctttg caagttcctc agcattctta acaaaagacg
     2461 tctcttttga catgtttaaa gtttaaacct cctgtgtgaa attattatcc gctcataatt
     2521 ccacacatta tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     2581 taactcacat taattgcgtt gcgctcactg ccaattgctt tccagtcggg aaacctgtcg
     2641 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     2701 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     2761 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     2821 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     2881 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     2941 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     3001 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     3061 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     3121 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     3181 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     3241 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     3301 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     3361 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     3421 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     3481 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     3541 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     3601 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     3661 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     3721 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     3781 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     3841 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     3901 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     3961 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     4021 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     4081 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     4141 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     4201 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     4261 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     4321 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     4381 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     4441 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     4501 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     4561 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     4621 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     4681 cgtatcacga ggcc
//
