LOCUS       sequence-415197-        4856 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Template for hlpA-mneongreen amplification associated with 
            spectinomycin resistance gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4856)
  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 4856)
  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: pJethlpAmNeonGreenSpc
FEATURES             Location/Qualifiers
     source          1..4856
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        305..323
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(join(2688..2694,2695..2712,2713..2718))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    2733..2754
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2874..2891)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3045..3633)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3125..3144)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3804..4595,4596..4664))
                     /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     4427..4446
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4665..4769)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4837..4855
                     /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 gccgctctcc ctatagtgag tcgtattggc
      361 cgccagatct tccggatggc tcgagttttt cagcaagatc atggcaaaca aacaagattt
      421 gatcgctaaa gtagcagaag ctacagaatt gactaagaaa gactcagcag cagcagttga
      481 agctgtattt gcagcagtag ctgactatct tgcagctggt gaaaaagttc aattgatcgg
      541 ttttggtaac tttgaagttc gtgagcgtgc agaacgtaaa ggtcgcaacc cacaaactgg
      601 taaagaaatg acaattgcag cttctaaagt accagcattc aaagctggta aagctcttaa
      661 agacgctgtt aaaggatccg gatctggtgg agaagctgca gctaaaggat cagtctctga
      721 agaagataat atggctagtt tgcctgctac ccacgaatta cacatttttg gatcaatcaa
      781 tggagtcgat tttgatatgg ttggtcaagg aaccggtaat ccaaatgatg gatacgaaga
      841 attgaatctt aagagtacta agggtgattt acagttttct ccatggattt tggtccctca
      901 tatcggatac ggttttcacc aatacttgcc atatcctgat ggaatgtctc cttttcaggc
      961 tgcaatggtt gatggatcag gttatcaagt ccatcgtaca atgcagtttg aagatggtgc
     1021 tagtcttacc gttaattatc gttacacata tgaaggatct cacattaaag gtgaagccca
     1081 agtaaaggga acaggttttc cagcagatgg acctgttatg actaattcac ttaccgccgc
     1141 tgattggtgt cgtagtaaaa agacttatcc aaatgataag acaatcatct ctacttttaa
     1201 gtggtcatac acaactggaa atggtaaacg ttatcgttca actgctcgta ccacatacac
     1261 ctttgcaaaa ccaatggcag ccaattacct taagaatcaa cctatgtacg tctttcgtaa
     1321 gacagaactt aagcattcta agactgaact taattttaag gaatggcaga aggcttttac
     1381 tgatgtaatg ggtatggatg aattgtataa ataataagaa ttccgaagtt atcgtaacgt
     1441 gactggcaag agatattttt aaaacaatga ataggtttac acttacttta gttttatgga
     1501 aatgaaagat catatcatat ataatctaga ataaaattaa ctaaaataat tattatctag
     1561 ataaaaaatt tagaagccaa tgaaatctat aaataaacta aattaagttt atttaattaa
     1621 caactatgga tataaaatag gtactaatca aaatagtgag gaggatatat ttgaatacat
     1681 acgaacaaat taataaagtg aaaaaaatac ttcggaaaca tttaaaaaat aaccttattg
     1741 gtacttacat gtttggatca ggagttgaga gtggactaaa accaaatagt gatcttgact
     1801 ttttagtcgt cgtatctgaa ccattgacag atcaaagtaa agaaatactt atacaaaaaa
     1861 ttagacctat ttcaaaaaaa ataggagata aaagcaactt acgatatatt gaattaacaa
     1921 ttattattca gcaagaaatg gtaccgtgga atcatcctcc caaacaagaa tttatttatg
     1981 gagaatggtt acaagagctt tatgaacaag gatacattcc tcagaaggaa ttaaattcag
     2041 atttaaccat aatgctttac caagcaaaac gaaaaaataa aagaatatac ggaaattatg
     2101 acttagagga attactacct gatattccat tttctgatgt gagaagagcc attatggatt
     2161 cgtcagagga attaatagat aattatcagg atgatgaaac caactctata ttaactttat
     2221 gccgtatgat tttaactatg gacacgggta aaatcatacc aaaagatatt gcgggaaatg
     2281 cagtggctga atcttctcca ttagaacata gggagagaat tttgttagca gttcgtagtt
     2341 atcttggaga gaatattgaa tggactaatg aaaatgtaaa tttaactata aactatttaa
     2401 ataacagatt aaaaaaatta taaaaaaatt gaaaaaatgg tggaaacact tttttaattt
     2461 ttttgtttta ttatttaata tttgggaaat attcattcta attggaagct tcaggaaact
     2521 gagacaggaa ttttattaaa aatttaaatt ttgaagaaag ttcagggtta atagcatcca
     2581 ttttttgctt tgcaagttcc tcagcattct taacaaaaga cgtctctttt gacatgttta
     2641 aagtttaaac ctcctgtgtg aaattattat ccgctcataa ttccacacat tatacgagcc
     2701 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     2761 ttgcgctcac tgccaattgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     2821 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     2881 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     2941 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     3001 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     3061 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     3121 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     3181 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     3241 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     3301 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     3361 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     3421 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     3481 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     3541 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     3601 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     3661 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     3721 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     3781 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     3841 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     3901 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     3961 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     4021 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     4081 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     4141 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     4201 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     4261 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     4321 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     4381 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     4441 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     4501 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     4561 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     4621 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     4681 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     4741 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     4801 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcc
//
