LOCUS       sequence-415189-        4816 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Template for hlpA-mkate2 amplification associated with spectinomycin
            resistance gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4816)
  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 4816)
  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: pJethlpAmKate2Spc
FEATURES             Location/Qualifiers
     source          1..4816
                     /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(2648..2654,2655..2672,2673..2678))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    2693..2714
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2834..2851)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3005..3593)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3085..3104)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3764..4555,4556..4624))
                     /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     4387..4406
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4625..4729)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4797..4815
                     /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 ttccgaagtt atcgtaacgt gactggcaag agatattttt aaaacaatga ataggtttac
     1441 acttacttta gttttatgga aatgaaagat catatcatat ataatctaga ataaaattaa
     1501 ctaaaataat tattatctag ataaaaaatt tagaagccaa tgaaatctat aaataaacta
     1561 aattaagttt atttaattaa caactatgga tataaaatag gtactaatca aaatagtgag
     1621 gaggatatat ttgaatacat acgaacaaat taataaagtg aaaaaaatac ttcggaaaca
     1681 tttaaaaaat aaccttattg gtacttacat gtttggatca ggagttgaga gtggactaaa
     1741 accaaatagt gatcttgact ttttagtcgt cgtatctgaa ccattgacag atcaaagtaa
     1801 agaaatactt atacaaaaaa ttagacctat ttcaaaaaaa ataggagata aaagcaactt
     1861 acgatatatt gaattaacaa ttattattca gcaagaaatg gtaccgtgga atcatcctcc
     1921 caaacaagaa tttatttatg gagaatggtt acaagagctt tatgaacaag gatacattcc
     1981 tcagaaggaa ttaaattcag atttaaccat aatgctttac caagcaaaac gaaaaaataa
     2041 aagaatatac ggaaattatg acttagagga attactacct gatattccat tttctgatgt
     2101 gagaagagcc attatggatt cgtcagagga attaatagat aattatcagg atgatgaaac
     2161 caactctata ttaactttat gccgtatgat tttaactatg gacacgggta aaatcatacc
     2221 aaaagatatt gcgggaaatg cagtggctga atcttctcca ttagaacata gggagagaat
     2281 tttgttagca gttcgtagtt atcttggaga gaatattgaa tggactaatg aaaatgtaaa
     2341 tttaactata aactatttaa ataacagatt aaaaaaatta taaaaaaatt gaaaaaatgg
     2401 tggaaacact tttttaattt ttttgtttta ttatttaata tttgggaaat attcattcta
     2461 attggaagct tcaggaaact gagacaggaa ttttattaaa aatttaaatt ttgaagaaag
     2521 ttcagggtta atagcatcca ttttttgctt tgcaagttcc tcagcattct taacaaaaga
     2581 cgtctctttt gacatgttta aagtttaaac ctcctgtgtg aaattattat ccgctcataa
     2641 ttccacacat tatacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     2701 gctaactcac attaattgcg ttgcgctcac tgccaattgc tttccagtcg ggaaacctgt
     2761 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     2821 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     2881 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     2941 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     3001 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     3061 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     3121 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     3181 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     3241 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     3301 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     3361 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     3421 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     3481 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     3541 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     3601 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     3661 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     3721 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     3781 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     3841 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     3901 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     3961 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     4021 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     4081 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     4141 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     4201 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     4261 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     4321 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     4381 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     4441 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     4501 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     4561 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     4621 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     4681 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     4741 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     4801 ggcgtatcac gaggcc
//
