LOCUS       sequence-415190-        4930 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Template for hlpA-mneongreen amplification associated with 
            chloramphenicol resistance gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4930)
  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 4930)
  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: pJethlpAmNeonGreenCm
FEATURES             Location/Qualifiers
     source          1..4930
                     /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(2762..2768,2769..2786,2787..2792))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    2807..2828
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2948..2965)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3119..3707)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3199..3218)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3878..4669,4670..4738))
                     /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     4501..4520
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4739..4843)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4911..4929
                     /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 aattccgtat tggtggaata acacgaatta ataaaggata tatttgtagt
     1441 agcaagtgta tttgttttat attctatgac cctatttgat ttacatgatt ctatatggta
     1501 taatggactt gcttataaat atagatgctt ggatagcagg aggttgcaag atgatgatgg
     1561 cagttaaatt agccttctaa aattaaataa taaaatttta ggaggctaat ttatgaaatt
     1621 taatttgata gatatagagg attggaatag aaagccatac tttgagcatt atttaaatgc
     1681 ggtaaggtgc acttacagta tgactgcaaa tttagagata actagtttac tgcgtgaaat
     1741 taaacttaag ggcctgaaac tgtaccctac gcttatttat atcatcacaa ctgtagttaa
     1801 tcgtcacaag gagttccgca ccagttttga tcaaaacggt aagttaggat actgggatag
     1861 tatgaaccca agttatacta tctttcataa ggataacgaa actttttcaa gtatttggac
     1921 agagtatgac gagaacttcc cacgtttcta ctataattac cttgaggata tcagaaacta
     1981 tagcgacgtt atgaatttca tgcctaaggc aggtcaacct gctaacacaa ttaatgtgtc
     2041 cagcattcct tgggtgaatt ttaccggatt caacttgaat atatataatg atgcaacata
     2101 tctaatccct atttttacta tgggtaagta ttttcagcag gataataaaa ttttattacc
     2161 tatgtctgta caggtgcatc atgctgtttg cgacggttac cataccagca gattttttaa
     2221 tgaggtacag gaattagtct caaattttga gacatggtta ggagaaaaat aatattaaaa
     2281 tgcttgtgct cgggagttat aaacaaagat cgaaaatatt gttaagatcg tgaggaagca
     2341 aagcgaccga agaagtatat gttccatcta tgcaaggact tcccgagaag gcgagtgaca
     2401 gcgaagaccg attggaagta gtctgagcta agtggagtaa tatattcatt cattatagaa
     2461 ccatcatttc aaaggattag agaaatctaa tcctttttct aggtattttt atcctaagaa
     2521 tttaatatga caatcacgag tttaattttt attttggtgc tatttatcct aagcagagca
     2581 agcttcagga aactgagaca ggaattttat taaaaattta aattttgaag aaagttcagg
     2641 gttaatagca tccatttttt gctttgcaag ttcctcagca ttcttaacaa aagacgtctc
     2701 ttttgacatg tttaaagttt aaacctcctg tgtgaaatta ttatccgctc ataattccac
     2761 acattatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     2821 tcacattaat tgcgttgcgc tcactgccaa ttgctttcca gtcgggaaac ctgtcgtgcc
     2881 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     2941 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     3001 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     3061 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     3121 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     3181 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     3241 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     3301 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     3361 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     3421 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     3481 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     3541 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     3601 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     3661 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     3721 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     3781 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     3841 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     3901 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     3961 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     4021 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     4081 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     4141 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     4201 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     4261 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     4321 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     4381 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     4441 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     4501 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     4561 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     4621 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     4681 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     4741 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     4801 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     4861 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     4921 tcacgaggcc
//
