LOCUS       sequence-210564-        5679 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 5679)
  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 5679)
  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: NK649
FEATURES             Location/Qualifiers
     source          1..5679
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(14..32)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        100..204
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(205..273,274..1065)
                     /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     complement(423..442)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1236..1824
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1725..1744
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1978..1995
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2112..2133
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2148..2153,2154..2171,2172..2178)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2186..2202
                     /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)."
     primer_bind     2191..2213
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2210..2226
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2210..2226
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5289..5305)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5298..5320)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5514..5533)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5633..5655
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
       61 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
      121 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
      181 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
      241 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
      301 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
      361 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
      421 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
      481 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
      541 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
      601 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
      661 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
      721 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
      781 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
      841 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
      901 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
      961 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     1021 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     1081 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     1141 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     1201 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     1261 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     1321 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     1381 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     1441 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     1501 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     1561 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     1621 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     1681 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     1741 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     1801 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     1861 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     1921 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     1981 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     2041 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     2101 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
     2161 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
     2221 tatgaccatg attacgccaa gcttgtaaca aactgccctg cacctcgtct gccacctcgg
     2281 ttgccacctt gtcctcgttc cccaccacct tgtcctcgtc cgctgccagc atcgcgatca
     2341 gcgtgcttgc gcttggcgct gctgctgccg gacttcccgt tatcggccat actttttgtg
     2401 gtgtttccag gcgagatgcg ctgttggcaa agcaaaatgg caaaacggcc tttgccacac
     2461 gccatcctcg cctcacctcg acgtgcggct ttcctaaaca ggaaacctga cgaaacgaat
     2521 tgagcgcatt ggcacacgtg tgctgtgcgc tttgtgtcgc tgcatgctat ccaccacgca
     2581 ccacccaatc agtccaactg tgacgtgtca cgcctaggcg aaaaagcggc gagagcgcca
     2641 aagcaaacgc caaacaaaaa ccaaagaaga gcaaacgcag cgaactgatg tgagacaagg
     2701 gggcggtgca atgcgtcact gaacgtgcaa aagccaaaaa caaccaagaa gcagctgcac
     2761 aacacaacaa caacaacaac aacaacaaca ccttgttgtc gcagctctac gtttggcatt
     2821 ttcctgttgt tctctcttgc gatcagtccg tgtctctttt ctcccccttt tcccagacca
     2881 caaccacaaa acaaccagcc atggcgacga tgaggacctt ggccccgtcg gggcgtcgct
     2941 tgactgcaac gtcccgtgtc ggctttgaca ccttgccgga tcagtatgtc aaccgcgaac
     3001 agcagcgagg cttccgcttc aacattgctt gcatcggcga gacggccatc ggcaagtctg
     3061 cgctgctgga ttccctgttt gacacggact ttgagcagaa gccacacaac cacgatgaag
     3121 agccacacgt cgagcacagc tgcaaggttg tgagcgaggg cggcgtggac ctgcagctca
     3181 ccatcacgtg ctccgtcggt tacggcgacc agattgaccg cacagacacc tacaagcctc
     3241 tcctcgacta cgtcgacgcc cagttcgagg agtacctgca ggaggagctg aagatgaacc
     3301 gacagctgca ttcatacacg gacacccgcg tgcacgcgtg cctgttcctc gtctctccca
     3361 cgggcacctg cctcaagtct atggatctcg ttgccctcaa gaagctgtgc gacaaggtca
     3421 atgtgattgt tgtcatcgcc aaggccgaca ccatcacaaa atccgagctc gagcgcttca
     3481 agcaaacgct tcgtgaggag tttgctgcca acggtttgca gctgttccag ccagaggttg
     3541 acacaaacca gtaccccctg gctgttgtcg gcagtcagga gaaggtcgtc attggtggcg
     3601 agagcgttcg tgtccgccaa tacccatggg gcatcgtcga agttgagaac gacgatcaca
     3661 gcgactttct tcgtctccgc aacctcctcc tcaggacaca ccttgaggtg ctgcggctgt
     3721 cgacacacac gacgttttac gagcgctatc gtcagcgccg gctcgttgat ctcgggtttg
     3781 cgccgtcggg ccgccacaca tccaccagcg gggcggacgg cgacgcagag gacgtgccgg
     3841 agtcgctgac ggaggtgtat gagcagaaac ggcggcagta caaagagctc atggcccgca
     3901 aggagcagca gctcaaggaa cagttcatgc agaaggtgaa cgagaaggag ggggagattc
     3961 gcaagattga gaaggaattg cgcgggaaac gcgatgagct cgtcaacatt caccgcagcg
     4021 aagaggccaa gatcaaggcc gctctccagg cgcttgcgcg ggaccgcgct gcgtggcagg
     4081 cggagtgcga ggcgcgacgc aaggaggaaa tgagcaagga ctcgaagaag aaggacaaga
     4141 aggacaagca ccgccacaag ggcggtagcg gtggctccat ggtctccaag ggcgaggagg
     4201 acaacatggc tatcatcaag gagttcatgc gcttcaaggt ccacatggag ggctccgtca
     4261 acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga
     4321 ccgccaagct caaggtgacg aagggcggcc ccctcccctt cgcctgggac atcctctccc
     4381 cccagttcat gtacggctcc aaggcttacg tcaagcaccc cgccgacatc cccgactacc
     4441 tcaagctctc cttccccgag ggcttcaagt gggagcgcgt catgaacttc gaggacggcg
     4501 gcgtcgtcac cgtcacccag gactcctctc ttcaggacgg cgagttcatc tacaaggtca
     4561 agctgcgcgg caccaacttc ccctccgacg gccccgtcat gcagaagaag acgatgggtt
     4621 gggaggcttc ctccgagcgc atgtaccccg aggacggcgc cctcaagggc gagatcaagc
     4681 agcgcctcaa gctcaaggac ggcggccact acgacgccga ggtcaagacc acctacaagg
     4741 ccaagaagcc cgtccagctc cccggcgcct acaacgtcaa catcaagctc gacatcacct
     4801 cccacaacga ggactacacc atcgtcgagc agtacgagcg tgccgagggc cgccacagca
     4861 cgggcggtat ggacgagctt tacaagtctt aagagctccc cccagcatta tcacgtctac
     4921 tgtgtgtatg ttgtcgctct ctgtttgacc tctctcttgt ctactatgtt gcctctctct
     4981 ctttcttgat tgcacccctt tcctcgctac gactactacg acttccgcta ccttgacttt
     5041 gttggcttgt tgccaccttt tggtcttgcg tttcgtctgc tcgtcactct tgtctctttt
     5101 tgacgacttg tgtgtgatga cggcgtgagg ctgtgactga ctgactctca acaacttcca
     5161 ataaaaaccc ctacatcgca cgtgactgac tgcacattta gtgaagggaa ggaaaagaag
     5221 tgaggtgttt tcatgtacat gaggcttgtc gttgctctag aggatccccg ggtaccgagc
     5281 tcgaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     5341 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     5401 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat
     5461 tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc
     5521 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
     5581 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     5641 tgcatgtgtc agaggttttc accgtcatca ccgaaacgc
//
