LOCUS       sequence-297995-        4362 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Reporter gene expression in S. rosetta.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4362)
  AUTHORS   Booth DS, Szmidt-Middleton H, King N
  TITLE     Transfection of choanoflagellates illuminates their cell biology and
            the ancestry of animal septins.
  JOURNAL   Mol Biol Cell. 2018 Dec 1;29(25):3026-3038. doi: 
            10.1091/mbc.E18-08-0514. Epub 2018 Oct 3.
  PUBMED    30281390
REFERENCE   2  (bases 1 to 4362)
  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"; doi: 
            "10.1091/mbc.E18-08-0514"; journalName: "Mol Biol Cell"; date: 
            "2018-12-1- 1"; volume: "29"; issue: "25"; pages: "3026-3038"
COMMENT     Sequence Label: NK624
FEATURES             Location/Qualifiers
     source          1..4362
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     95..114
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     308..330
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     322..339
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     323..339
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2085..2101)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2085..2101)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2098..2120)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2109..2125
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2133..2139,2140..2157,2158..2163))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2178..2199
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2316..2333)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2487..3075)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2567..2586)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3246..4037,4038..4106))
                     /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     3869..3888
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4107..4211)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4279..4297
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4335..4357)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
       61 ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga
      121 gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
      181 cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
      241 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca
      301 gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattcgagct cggtacccgg
      361 ggatcctcta gacgggcacc gctcattgat cgtcacagtt atacattggg gggaaggata
      421 gttacactgg catagtgggt ttattgcggg gcccaagggc tcggcatgac gtgcacaaga
      481 agcagaatca actcacaaca agcgcacgtg gcacgcgtcg acacgaggac aacagcaaaa
      541 gaggcaacaa ggtggcaatg accaacgagg cgcttagagc tcgtcgtggc taccgccgga
      601 gccaccagac ttgtaaagct cgtccatacc gcccgtgctg tggcggccct cggcacgctc
      661 gtactgctcg acgatggtgt agtcctcgtt gtgggaggtg atgtcgagct tgatgttgac
      721 gttgtaggcg ccggggagct ggacgggctt cttggccttg taggtggtct tgacctcggc
      781 gtcgtagtgg ccgccgtcct tgagcttgag gcgctgcttg atctcgccct tgagggcgcc
      841 gtcctcgggg tacatgcgct cggaggaagc ctcccaaccc atcgtcttct tctgcatgac
      901 ggggccgtcg gaggggaagt tggtgccgcg cagcttgacc ttgtagatga actcgccgtc
      961 ctgaagagag gagtcctggg tgacggtgac gacgccgccg tcctcgaagt tcatgacgcg
     1021 ctcccacttg aagccctcgg ggaaggagag cttgaggtag tcggggatgt cggcggggtg
     1081 cttgacgtaa gccttggagc cgtacatgaa ctggggggag aggatgtccc aggcgaaggg
     1141 gagggggccg cccttcgtca ccttgagctt ggcggtctgg gtgccctcgt aggggcggcc
     1201 ctcgccctcg ccctcgatct cgaactcgtg gccgttgacg gagccctcca tgtggacctt
     1261 gaagcgcatg aactccttga tgatagccat gttgtcctcc tcgcccttgg agaccatgga
     1321 gccaccgcta ccgcctgcaa catcaacggg cgcagctgtg gccacgcagg ctgccatcac
     1381 gatggcggcg gtggtcaaca ccagcagcat ggctggttgt tttgtggttg tggtctggga
     1441 aaagggggag aaaagagaca cggactgatc gcaagagaga acaacaggaa aatgccaaac
     1501 gtagagctgc gacaacaagg tgttgttgtt gttgttgttg ttgttgtgtt gtgcagctgc
     1561 ttcttggttg tttttggctt ttgcacgttc agtgacgcat tgcaccgccc ccttgtctca
     1621 catcagttcg ctgcgtttgc tcttctttgg tttttgtttg gcgtttgctt tggcgctctc
     1681 gccgcttttt cgcctaggcg tgacacgtca cagttggact gattgggtgg tgcgtggtgg
     1741 atagcatgca gcgacacaaa gcgcacagca cacgtgtgcc aatgcgctca attcgtttcg
     1801 tcaggtttcc tgtttaggaa agccgcacgt cgaggtgagg cgaggatggc gtgtggcaaa
     1861 ggccgttttg ccattttgct ttgccaacag cgcatctcgc ctggaaacac cacaaaaagt
     1921 atggccgata acgggaagtc cggcagcagc agcgccaagc gcaagcacgc tgatcgcgat
     1981 gctggcagcg gacgaggaca aggtggtggg gaacgaggac aaggtggcaa ccgaggtggc
     2041 agacgaggtg cagggcagtt tgttacaagc ttggcgtaat catggtcata gctgtttcct
     2101 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     2161 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     2221 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     2281 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     2341 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     2401 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     2461 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     2521 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     2581 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     2641 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     2701 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     2761 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     2821 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     2881 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     2941 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     3001 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     3061 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     3121 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     3181 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     3241 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     3301 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     3361 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     3421 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     3481 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     3541 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     3601 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     3661 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     3721 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     3781 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     3841 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     3901 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     3961 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     4021 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     4081 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     4141 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     4201 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     4261 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt
     4321 gatgacggtg aaaacctctg acacatgcag ctcccggaga cg
//
