LOCUS       sequence-205763-        5956 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 5956)
  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 5956)
  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: NK639
FEATURES             Location/Qualifiers
     source          1..5956
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        96..200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(201..269,270..1061)
                     /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(419..438)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1232..1820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1721..1740
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1974..1991
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2108..2129
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2144..2149,2150..2167,2168..2174)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2182..2198
                     /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     2187..2209
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2206..2222
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2206..2222
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5562..5578)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5571..5593)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5787..5806)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5906..5928
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacatc ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2101 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     2161 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     2221 accatgatta cgccaagctt gtaacgactg cagtcttgtt gtcgtttgtc gtcgtcgttc
     2281 tctgtctccc acccaacttt tgccaattgc aacagtgact tctgtacaat ttgcaacaga
     2341 taccaacgtg tcgagtcagc acaaatggcg aggaaggttc gcgtgcattg cttgcgcgcc
     2401 tctttgtcat gggcgcacgc tgatttgtgt gtgatttgca ccaaatttcc tgccgacaca
     2461 acttgggccc gtaccacacg acccgtttct ggacccgttt ctggcccaga aacgcgaatg
     2521 ttgcgaaaga tgcgaccacg tgctcgctgt cgacaacaca aactcccgca caatccccat
     2581 gaaaaaaaaa gtgcttgctg ggcttgtgtc tggcctgtgt gcgtggcgtg gccgcgtgtg
     2641 gcttcgttct tgcttctgct cactcagcct gttacacgca gagtggctgc gagaaacaag
     2701 gagaggggtt tgcggcattg gcgcatgtgc atctgccttt ttcattgagt cagtctgcag
     2761 ctcaagaggt tggtttcgtc gacggatatg agtgagcttc tatggatgat gagaggtggt
     2821 ggtggttgta tggttggtgg ttggtggttg gtgcaaggcg agccacgcga agagcgacaa
     2881 ggatgaggga gaggagagga gactaggaaa gaggtgaagg gaagggaagg gcgagcaatg
     2941 acgacgagat aaaagacagc caccaccgca cccgcacaaa agcattgctc gcacgcatgg
     3001 tcgtgtgtta tgtttcacac caccaacata accaatacaa gccatcacca atacaagcca
     3061 gcgctcaagc acacacgcac gcacgcacac acaatacaca cgcacgtgaa acgttcacat
     3121 tgtccgctca caaatgcctt tcccttctcg tcgctttcct tgcctttttt tcttcccctc
     3181 ctgacttcct tcctgccttc cttccttcct ccccttctcc gtccaccaac accatttatt
     3241 tctctgctct ttttcctcgt gccacgtgag cagtcttttc gcgtcgtctt cttctttgac
     3301 cccgtttacc gtcaacacca tcaacaacca gcaacatggc gacgatgagg accttggccc
     3361 cgtcggggcg tcgcttgact gcaacgtccc gtgtcggctt tgacaccttg ccggatcagt
     3421 atgtcaaccg cgaacagcag cgaggcttcc gcttcaacat tgcttgcatc ggcgagacgg
     3481 ccatcggcaa gtctgcgctg ctggattccc tgtttgacac ggactttgag cagaagccac
     3541 acaaccacga tgaagagcca cacgtcgagc acagctgcaa ggttgtgagc gagggcggcg
     3601 tggacctgca gctcaccatc acgtgctccg tcggttacgg cgaccagatt gaccgcacag
     3661 acacctacaa gcctctcctc gactacgtcg acgcccagtt cgaggagtac ctgcaggagg
     3721 agctgaagat gaaccgacag ctgcattcat acacggacac ccgcgtgcac gcgtgcctgt
     3781 tcctcgtctc tcccacgggc acctgcctca agtccatgga tctcgttgcc ctcaagaagc
     3841 tgtgcgacaa ggtcaatgtg attgttgtca tcgccaaggc cgacaccatc accaaatccg
     3901 agctcgagcg cttcaagcaa acgcttcgtg aggagtttgc tgccaacggt ttgcagctgt
     3961 tccagccaga ggttgacaca aaccagtacc ccctggctgt tgtcggcagt caggagaagg
     4021 tcgtcattgg tggcgagagc gttcgtgtcc gccaataccc atggggcatc gtcgaagttg
     4081 agaacgacga tcacagcgac tttcttcgtc tccgcaacct cctcctcagg acacaccttg
     4141 aggtgctgcg gctgtcgaca cacacgacgt tttacgagcg ctatcgtcag cgccggctcg
     4201 ttgatctcgg gtttgcgccg tcgggccgcc acacatccac cagcggggcg gacggcgacg
     4261 cagaggacgt gccggagtcg ctgacggagg tgtatgagca gaaacggcgg cagtacaaag
     4321 agctcatggc ccgcaaggag cagcagctca aggaacagtt catgcagaag gtgaacgaga
     4381 aggaggggga gattcgcaag attgagaagg aattgcgcgg gaaacgcgat gagctcgtca
     4441 acattcaccg cagcgaagag gccaagatca aggccgctct ccaggcgctt gcgcgggacc
     4501 gcgctgcgtg gcaggcggag tgcgaggcgc gacgcaagga ggaaatgagc aaggactcga
     4561 agaagaagga caagaaggac aagcaccgcc acaagtctgg cggtagcggt ggctccgtct
     4621 ccaagggcga ggagaccacg atgggcgtca tcaagcccga catgaagatc aagctcaaga
     4681 tggagggcaa cgtcaacggc cacgccttcg tcatcgaggg cgagggcgag ggcaagccct
     4741 acgacggcac caacaccatc aacctcgagg tcaaggaggg cgcccccctc cccttctcct
     4801 acgacatcct caccaccgcc ttcgcctacg gcaaccgcgc cttcaccaag taccccgacg
     4861 acatccccaa ctacttcaag cagtccttcc ccgagggcta ctcctgggag cgcaccatga
     4921 ccttcgagga caagggcatc gtcaaggtca agtccgacat ctctatggag gaggactcct
     4981 tcatctacga gatccacctc aagggcgaga acttcccccc caacggcccc gtcatgcaga
     5041 agaagaccac cggctgggac gcctccaccg agcgcatgta cgtccgcgac ggcgtcctca
     5101 agggcgacgt gaagcacaag ctcctcctcg agggcggcgg ccaccaccgc gtcgacttca
     5161 agaccatcta ccgcgccaag aaggccgtca agctccccga ctaccacttc gtcgaccacc
     5221 gcatcgagat cctcaaccac gacaaggact acaacaaggt caccgtctac gagtccgccg
     5281 tcgcccgcaa ctccaccgac ggcatggacg agctgtacaa ggcttaagcg cctcgttggt
     5341 cattgccacc ttgttgcctc ttttgctgtt gtcctcgtgt cgacgcgtgc cacgtgcgct
     5401 tgttgtgagt tgattctgct tcttgtgcac gtcatgccga gcccttgggc cccgcaataa
     5461 acccactatg ccagtgtaac tatccttccc cccaatgtat aactgtgacg atcaatgagc
     5521 ggtgcccgtc tagaggatcc ccgggtaccg agctcgaatt cactggccgt cgttttacaa
     5581 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
     5641 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     5701 agcctgaatg gcgaatggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     5761 tcacaccgca tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     5821 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     5881 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     5941 tcaccgaaac gcgcga
//
