LOCUS       sequence-210428-        4959 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 4959)
  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 4959)
  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: NK597
FEATURES             Location/Qualifiers
     source          1..4959
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(5..23)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        91..195
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(196..264,265..1056)
                     /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(414..433)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1227..1815
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1716..1735
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1969..1986
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2103..2124
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2139..2144,2145..2162,2163..2169)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2177..2193
                     /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     2182..2204
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2201..2217
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2201..2217
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4560..4577)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4560..4576)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4569..4591)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4785..4804)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4904..4926
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
       61 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
      121 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
      181 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
      241 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
      301 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
      361 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
      421 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
      481 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
      541 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
      601 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
      661 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
      721 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
      781 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
      841 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
      901 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
      961 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     1021 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     1081 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     1141 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     1201 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     1261 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     1321 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     1381 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     1441 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     1501 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     1561 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     1621 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     1681 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     1741 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     1801 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     1861 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     1921 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     1981 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     2041 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     2101 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
     2161 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat
     2221 gattacgcca agcttgtaac aaactgccct gcacctcgtc tgccacctcg gttgccacct
     2281 tgtcctcgtt ccccaccacc ttgtcctcgt ccgctgccag catcgcgatc agcgtgcttg
     2341 cgcttggcgc tgctgctgcc ggacttcccg ttatcggcca tactttttgt ggtgtttcca
     2401 ggcgagatgc gctgttggca aagcaaaatg gcaaaacggc ctttgccaca cgccatcctc
     2461 gcctcacctc gacgtgcggc tttcctaaac aggaaacctg acgaaacgaa ttgagcgcat
     2521 tggcacacgt gtgctgtgcg ctttgtgtcg ctgcatgcta tccaccacgc accacccaat
     2581 cagtccaact gtgacgtgtc acgcctaggc gaaaaagcgg cgagagcgcc aaagcaaacg
     2641 ccaaacaaaa accaaagaag agcaaacgca gcgaactgat gtgagacaag ggggcggtgc
     2701 aatgcgtcac tgaacgtgca aaagccaaaa acaaccaaga agcagctgca caacacaaca
     2761 acaacaacaa caacaacaac accttgttgt cgcagctcta cgtttggcat tttcctgttg
     2821 ttctctcttg cgatcagtcc gtgtctcttt tctccccctt ttcccagacc acaaccacaa
     2881 aacaaccagc catggtctcc aagggcgagg aggtgatcaa ggagtttatg cgctttaagg
     2941 tccgtatgga gggctctatg aacggccacg agtttgagat cgagggtgag ggcgagggtc
     3001 gcccttacga gggcacgcag accgctaagc tcaaggtgac caagggtggc cctctccctt
     3061 tcgcttggga tatcctgtcc cctcagttca tgtacggttc caaggcttac gtcaagcacc
     3121 ctgccgatat ccctgactac aagaagctgt cctttcccga gggtttcaag tgggagcgtg
     3181 tcatgaactt tgaggacggt ggcctggtca cggtcacgca ggactcttcc ctgcaggacg
     3241 gtacgctgat ctacaaggtg aagatgcgcg gtacgaactt tccccctgac ggtcccgtga
     3301 tgcagaagaa gacgatgggt tgggaggcct ctaccgagcg cctgtacccc cgcgatggcg
     3361 tgctcaaggg tgagatccac caggctctca agctgaagga cggtggccac tacctcgtgg
     3421 agttcaagac gatctacatg gctaagaagc ccgtgcagct ccctggctac tattacgtgg
     3481 acacgaagct cgacattacc tctcacaacg aggactacac gatcgttgag cagtacgagc
     3541 gttctgaggg ccgtcaccac ctctttctcg gtcacggtac gggttctact ggtgccggct
     3601 cctctggcac ggcctcttcc gaggataaca acatggccgt cattaaggag ttcatgcgtt
     3661 tcaaggtgcg catggagggt tccatgaacg gtcacgagtt cgagattgag ggcgagggtg
     3721 agggccgtcc ctacgagggt acgcagacgg ccaagctgaa ggtcacgaag ggcggtcccc
     3781 tgccctttgc ctgggatatt ctctctcccc agtttatgta cggctctaag gcctacgtga
     3841 agcaccccgc tgacattccc gattacaaga agctctcttt ccctgagggc tttaagtggg
     3901 agcgcgtgat gaacttcgag gatggcggtc tcgtgaccgt gacccaggat tcctctctgc
     3961 aggatggcac gctcatttac aaggtcaaga tgcgtggcac gaacttccct cccgatggcc
     4021 ctgtcatgca gaagaagacg atgggctggg aggcttccac ggagcgtctc taccctcgtg
     4081 acggtgtcct gaagggcgag attcaccagg ccctgaagct caaggatggc ggtcactacc
     4141 tggtcgagtt taagacgatt tacatggcca agaagcctgt ccagctgccc ggttactact
     4201 acgtcgatac caagctggat atcacgtccc acaacgagga ttacaccatt gtcgagcagt
     4261 acgagcgttc cgagggtcgc caccacctgt tcctgtacgg catggacgag ctgtacaagg
     4321 cttaagcgcc tcgttggtca ttgccacctt gttgcctctt ttgctgttgt cctcgtgtcg
     4381 acgcgtgcca cgtgcgcttg ttgtgagttg attctgcttc ttgtgcacgt catgccgagc
     4441 ccttgggccc cgcaataaac ccactatgcc agtgtaacta tccttccccc caatgtataa
     4501 ctgtgacgat caatgagcgg tgcccgtcta gaggatcccc gggtaccgag ctcgaattca
     4561 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     4621 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     4681 ccttcccaac agttgcgcag cctgaatggc gaatggcgcc tgatgcggta ttttctcctt
     4741 acgcatctgt gcggtatttc acaccgcata tggtgcactc tcagtacaat ctgctctgat
     4801 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     4861 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     4921 cagaggtttt caccgtcatc accgaaacgc gcgagacga
//
