LOCUS       sequence-316093-        4401 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 4401)
  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 4401)
  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: NK648
FEATURES             Location/Qualifiers
     source          1..4401
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..46)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(200..788)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(280..299)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(959..1750,1751..1819))
                     /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     1582..1601
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1820..1924)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1992..2010
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2048..2070)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2170..2189
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2383..2405
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2397..2414
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2398..2414
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4199..4215)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4199..4215)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4212..4234)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4223..4239
                     /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(4247..4253,4254..4271,4272..4277))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4292..4313
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
       61 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
      121 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
      181 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
      241 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
      301 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
      361 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
      421 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
      481 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
      541 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
      601 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
      661 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
      721 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
      781 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
      841 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
      901 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
      961 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     1021 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     1081 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     1141 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     1201 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     1261 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     1321 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     1381 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     1441 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     1501 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     1561 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     1621 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     1681 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     1741 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     1801 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     1861 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     1921 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     1981 taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg
     2041 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     2101 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
     2161 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac
     2221 cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca
     2281 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
     2341 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
     2401 aaacgacggc cagtgaattc gagctcggta cccggggatc ctctagagca acgacaagcc
     2461 tcatgtacat gaaaacacct cacttctttt ccttcccttc actaaatgtg cagtcagtca
     2521 cgtgcgatgt aggggttttt attggaagtt gttgagagtc agtcagtcac agcctcacgc
     2581 cgtcatcaca cacaagtcgt caaaaagaga caagagtgac gagcagacga aacgcaagac
     2641 caaaaggtgg caacaagcca acaaagtcaa ggtagcggaa gtcgtagtag tcgtagcgag
     2701 gaaaggggtg caatcaagaa agagagagag gcaacatagt agacaagaga gaggtcaaac
     2761 agagagcgac aacatacaca cagtagacgt gataatgctg gggggagctc ttaagacttg
     2821 taaagctcgt ccataccgcc cgtgctgtgg cggccctcgg cacgctcgta ctgctcgacg
     2881 atggtgtagt cctcgttgtg ggaggtgatg tcgagcttga tgttgacgtt gtaggcgccg
     2941 gggagctgga cgggcttctt ggccttgtag gtggtcttga cctcggcgtc gtagtggccg
     3001 ccgtccttga gcttgaggcg ctgcttgatc tcgcccttga gggcgccgtc ctcggggtac
     3061 atgcgctcgg aggaagcctc ccaacccatc gtcttcttct gcatgacggg gccgtcggag
     3121 gggaagttgg tgccgcgcag cttgaccttg tagatgaact cgccgtcctg aagagaggag
     3181 tcctgggtga cggtgacgac gccgccgtcc tcgaagttca tgacgcgctc ccacttgaag
     3241 ccctcgggga aggagagctt gaggtagtcg gggatgtcgg cggggtgctt gacgtaagcc
     3301 ttggagccgt acatgaactg gggggagagg atgtcccagg cgaaggggag ggggccgccc
     3361 ttcgtcacct tgagcttggc ggtctgggtg ccctcgtagg ggcggccctc gccctcgccc
     3421 tcgatctcga actcgtggcc gttgacggag ccctccatgt ggaccttgaa gcgcatgaac
     3481 tccttgatga tagccatgtt gtcctcctcg cccttggaga ccatggctgg ttgttttgtg
     3541 gttgtggtct gggaaaaggg ggagaaaaga gacacggact gatcgcaaga gagaacaaca
     3601 ggaaaatgcc aaacgtagag ctgcgacaac aaggtgttgt tgttgttgtt gttgttgttg
     3661 tgttgtgcag ctgcttcttg gttgtttttg gcttttgcac gttcagtgac gcattgcacc
     3721 gcccccttgt ctcacatcag ttcgctgcgt ttgctcttct ttggtttttg tttggcgttt
     3781 gctttggcgc tctcgccgct ttttcgccta ggcgtgacac gtcacagttg gactgattgg
     3841 gtggtgcgtg gtggatagca tgcagcgaca caaagcgcac agcacacgtg tgccaatgcg
     3901 ctcaattcgt ttcgtcaggt ttcctgttta ggaaagccgc acgtcgaggt gaggcgagga
     3961 tggcgtgtgg caaaggccgt tttgccattt tgctttgcca acagcgcatc tcgcctggaa
     4021 acaccacaaa aagtatggcc gataacggga agtccggcag cagcagcgcc aagcgcaagc
     4081 acgctgatcg cgatgctggc agcggacgag gacaaggtgg tggggaacga ggacaaggtg
     4141 gcaaccgagg tggcagacga ggtgcagggc agtttgttac aagcttggcg taatcatggt
     4201 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     4261 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     4321 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     4381 gccaacgcgc ggggagaggc g
//
