LOCUS       sequence-415330-        4876 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Template for hlpA-YFP amplification associated with spectinomycin 
            resistance gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4876)
  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 4876)
  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: pJethlpAYFPSpc
FEATURES             Location/Qualifiers
     source          1..4876
                     /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(2708..2714,2715..2732,2733..2738))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    2753..2774
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2894..2911)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3065..3653)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3145..3164)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3824..4615,4616..4684))
                     /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     4447..4466
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4685..4789)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4857..4875
                     /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 cagctaaatc tagttctaga ggatctggtg gagaagctgc agctaaagct
      721 ggatcaaaag gagaagaact ttttacaggt gtagtaccta tcttggttga attggatggt
      781 gatgttaacg gtcacaaatt ttctgtacgt ggtgaaggtg aaggtgatgc aactaacggt
      841 aaattgacac ttaaattgat ttgtacaact ggaaaacttc ctgttccttg gcctactctt
      901 gttacaacat tgggttatgg acttatgtgt tttgctcgtt atcctgatca tatgaaacgt
      961 cacgattttt ttaaatctgc tatgccagaa ggttatgtac aagaacgtac aatttcattt
     1021 aaagatgacg gaacatataa aacacgtgct gaagtaaaat tcgaaggtga cactcttgtt
     1081 aatcgtatcg aattgaaagg aatcgatttc aaagaagatg gtaacatttt gggacacaaa
     1141 cttgaataca acttcaactc tcataatgtt tatatcacag ctgacaaaca aaaaaacggt
     1201 attaaagcta attttaaaat tcgtcacaat gttgaagatg gaggagttca attggctgat
     1261 cattatcaac aaaatacacc aatcggagac ggaccagtat tgcttccaga taaccactac
     1321 ctttcttatc aatcaaaact ttcaaaagat cctaacgaaa aacgtgacca tatggtactt
     1381 cttgaatttg ttacagcagc aggtatcact cacggtatgg acgaacttta taaataagaa
     1441 ttccgaagtt atcgtaacgt gactggcaag agatattttt aaaacaatga ataggtttac
     1501 acttacttta gttttatgga aatgaaagat catatcatat ataatctaga ataaaattaa
     1561 ctaaaataat tattatctag ataaaaaatt tagaagccaa tgaaatctat aaataaacta
     1621 aattaagttt atttaattaa caactatgga tataaaatag gtactaatca aaatagtgag
     1681 gaggatatat ttgaatacat acgaacaaat taataaagtg aaaaaaatac ttcggaaaca
     1741 tttaaaaaat aaccttattg gtacttacat gtttggatca ggagttgaga gtggactaaa
     1801 accaaatagt gatcttgact ttttagtcgt cgtatctgaa ccattgacag atcaaagtaa
     1861 agaaatactt atacaaaaaa ttagacctat ttcaaaaaaa ataggagata aaagcaactt
     1921 acgatatatt gaattaacaa ttattattca gcaagaaatg gtaccgtgga atcatcctcc
     1981 caaacaagaa tttatttatg gagaatggtt acaagagctt tatgaacaag gatacattcc
     2041 tcagaaggaa ttaaattcag atttaaccat aatgctttac caagcaaaac gaaaaaataa
     2101 aagaatatac ggaaattatg acttagagga attactacct gatattccat tttctgatgt
     2161 gagaagagcc attatggatt cgtcagagga attaatagat aattatcagg atgatgaaac
     2221 caactctata ttaactttat gccgtatgat tttaactatg gacacgggta aaatcatacc
     2281 aaaagatatt gcgggaaatg cagtggctga atcttctcca ttagaacata gggagagaat
     2341 tttgttagca gttcgtagtt atcttggaga gaatattgaa tggactaatg aaaatgtaaa
     2401 tttaactata aactatttaa ataacagatt aaaaaaatta taaaaaaatt gaaaaaatgg
     2461 tggaaacact tttttaattt ttttgtttta ttatttaata tttgggaaat attcattcta
     2521 attggaagct tcaggaaact gagacaggaa ttttattaaa aatttaaatt ttgaagaaag
     2581 ttcagggtta atagcatcca ttttttgctt tgcaagttcc tcagcattct taacaaaaga
     2641 cgtctctttt gacatgttta aagtttaaac ctcctgtgtg aaattattat ccgctcataa
     2701 ttccacacat tatacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     2761 gctaactcac attaattgcg ttgcgctcac tgccaattgc tttccagtcg ggaaacctgt
     2821 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     2881 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     2941 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     3001 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     3061 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     3121 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     3181 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     3241 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     3301 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     3361 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     3421 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     3481 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     3541 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     3601 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     3661 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     3721 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     3781 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     3841 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     3901 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     3961 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     4021 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     4081 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     4141 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     4201 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     4261 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     4321 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     4381 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     4441 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     4501 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     4561 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     4621 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     4681 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     4741 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     4801 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     4861 ggcgtatcac gaggcc
//
