LOCUS       sequence-210546-        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: NK640
FEATURES             Location/Qualifiers
     source          1..5956
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(11..29)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        97..201
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(202..270,271..1062)
                     /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(420..439)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1233..1821
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1722..1741
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1975..1992
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2109..2130
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2145..2150,2151..2168,2169..2175)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2183..2199
                     /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     2188..2210
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2207..2223
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2207..2223
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5563..5579)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5572..5594)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5788..5807)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5907..5929
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
       61 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
      121 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
      181 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
      241 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
      301 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
      361 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
      421 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
      481 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
      541 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
      601 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
      661 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
      721 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
      781 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
      841 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
      901 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
      961 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     1021 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     1081 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     1141 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     1201 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     1261 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     1321 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     1381 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     1441 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     1501 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     1561 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     1621 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     1681 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     1741 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     1801 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     1861 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     1921 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     1981 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     2041 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     2101 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     2161 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     2221 gaccatgatt acgccaagct tgtaacgact gcagtcttgt tgtcgtttgt cgtcgtcgtt
     2281 ctctgtctcc cacccaactt ttgccaattg caacagtgac ttctgtacaa tttgcaacag
     2341 ataccaacgt gtcgagtcag cacaaatggc gaggaaggtt cgcgtgcatt gcttgcgcgc
     2401 ctctttgtca tgggcgcacg ctgatttgtg tgtgatttgc accaaatttc ctgccgacac
     2461 aacttgggcc cgtaccacac gacccgtttc tggacccgtt tctggcccag aaacgcgaat
     2521 gttgcgaaag atgcgaccac gtgctcgctg tcgacaacac aaactcccgc acaatcccca
     2581 tgaaaaaaaa agtgcttgct gggcttgtgt ctggcctgtg tgcgtggcgt ggccgcgtgt
     2641 ggcttcgttc ttgcttctgc tcactcagcc tgttacacgc agagtggctg cgagaaacaa
     2701 ggagaggggt ttgcggcatt ggcgcatgtg catctgcctt tttcattgag tcagtctgca
     2761 gctcaagagg ttggtttcgt cgacggatat gagtgagctt ctatggatga tgagaggtgg
     2821 tggtggttgt atggttggtg gttggtggtt ggtgcaaggc gagccacgcg aagagcgaca
     2881 aggatgaggg agaggagagg agactaggaa agaggtgaag ggaagggaag ggcgagcaat
     2941 gacgacgaga taaaagacag ccaccaccgc acccgcacaa aagcattgct cgcacgcatg
     3001 gtcgtgtgtt atgtttcaca ccaccaacat aaccaataca agccatcacc aatacaagcc
     3061 agcgctcaag cacacacgca cgcacgcaca cacaatacac acgcacgtga aacgttcaca
     3121 ttgtccgctc acaaatgcct ttcccttctc gtcgctttcc ttgccttttt ttcttcccct
     3181 cctgacttcc ttcctgcctt ccttccttcc tccccttctc cgtccaccaa caccatttat
     3241 ttctctgctc tttttcctcg tgccacgtga gcagtctttt cgcgtcgtct tcttctttga
     3301 ccccgtttac cgtcaacacc atcaacaacc agcaacatgg tctccaaggg cgaggagacc
     3361 acgatgggcg tcatcaagcc cgacatgaag atcaagctca agatggaggg caacgtcaac
     3421 ggccacgcct tcgtcatcga gggcgagggc gagggcaagc cctacgacgg caccaacacc
     3481 atcaacctcg aggtcaagga gggcgccccc ctccccttct cctacgacat cctcaccacc
     3541 gccttcgcct acggcaaccg cgccttcacc aagtaccccg acgacatccc caactacttc
     3601 aagcagtcct tccccgaggg ctactcctgg gagcgcacca tgaccttcga ggacaagggc
     3661 atcgtcaagg tcaagtccga catctctatg gaggaggact ccttcatcta cgagatccac
     3721 ctcaagggcg agaacttccc ccccaacggc cccgtcatgc agaagaagac caccggctgg
     3781 gacgcctcca ccgagcgcat gtacgtccgc gacggcgtcc tcaagggcga cgtgaagcac
     3841 aagctcctcc tcgagggcgg cggccaccac cgcgtcgact tcaagaccat ctaccgcgcc
     3901 aagaaggccg tcaagctccc cgactaccac ttcgtcgacc accgcatcga gatcctcaac
     3961 cacaacaagg actacaacaa ggtcaccgtc tacgagtccg ccgtcgcccg caactccacc
     4021 gacggcatgg acgagctgta caaggcttct ggcggtagcg gtggctccgc gacgatgagg
     4081 accttggccc cgtcggggcg tcgcttgact gcaacgtccc gtgtcggctt tgacaccttg
     4141 ccggatcagt atgtcaaccg cgaacagcag cgaggcttcc gcttcaacat tgcttgcatc
     4201 ggcgagacgg ccatcggcaa gtctgcgctg ctggattccc tgtttgacac ggactttgag
     4261 cagaagccac acaaccacga tgaagagcca cacgtcgagc acagctgcaa ggttgtgagc
     4321 gagggcggcg tggacctgca gctcaccatc acgtgctccg tcggttacgg cgaccagatt
     4381 gaccgcacag acacctacaa gcctctcctc gactacgtcg acgcccagtt cgaggagtac
     4441 ctgcaggagg agctgaagat gaaccgacag ctgcattcat acacggacac ccgcgtgcac
     4501 gcgtgcctgt tcctcgtctc tcccacgggc acctgcctca agtccatgga tctcgttgcc
     4561 ctcaagaagc tgtgcgacaa ggtcaatgtg attgttgtca tcgccaaggc cgacaccatc
     4621 acaaaatccg agctcgagcg cttcaagcaa acgcttcgtg aggagtttgc tgccaacggt
     4681 ttgcagctgt tccagccaga ggttgacaca aaccagtacc ccctggctgt tgtcggcagt
     4741 caggagaagg tcgtcattgg tggcgagagc gttcgtgtcc gccaataccc atggggcatc
     4801 gtcgaagttg agaacgacga tcacagcgac tttcttcgtc tccgcaacct cctcctcagg
     4861 acacaccttg aggtgctgcg gctgtcgaca cacacgacgt tttacgagcg ctatcgtcag
     4921 cgccggctcg ttgatctcgg gtttgcgccg tcgggccgcc acacatccac cagcggggcg
     4981 gacggcgacg cagaggacgt gccggagtcg ctgacggagg tgtatgagca gaaacggcgg
     5041 cagtacaaag agctcatggc ccgcaaggag cagcagctca aggaacagtt catgcagaag
     5101 gtgaacgaga aggaggggga gattcgcaag attgagaagg aattgcgcgg gaaacgcgat
     5161 gagctcgtca acattcaccg cagcgaagag gccaagatca aggccgctct ccaggcgctt
     5221 gcgcgggacc gcgctgcgtg gcaggcggag tgcgaggcgc gacgcaagga ggaaatgagc
     5281 aaggactcga agaagaagga caagaaggac aagcaccgcc acaagtaagc gcctcgttgg
     5341 tcattgccac cttgttgcct cttttgctgt tgtcctcgtg tcgacgcgtg ccacgtgcgc
     5401 ttgttgtgag ttgattctgc ttcttgtgca cgtcatgccg agcccttggg ccccgcaata
     5461 aacccactat gccagtgtaa ctatccttcc ccccaatgta taactgtgac gatcaatgag
     5521 cggtgcccgt ctagaggatc cccgggtacc gagctcgaat tcactggccg tcgttttaca
     5581 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
     5641 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     5701 cagcctgaat ggcgaatggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     5761 ttcacaccgc atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     5821 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     5881 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     5941 atcaccgaaa cgcgcg
//
