LOCUS       sequence-210565-        5617 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 5617)
  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 5617)
  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: NK651
FEATURES             Location/Qualifiers
     source          1..5617
                     /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(5218..5235)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5218..5234)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5227..5249)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5443..5462)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5562..5584
                     /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 gactgcagtc ttgttgtcgt ttgtcgtcgt cgttctctgt
     2281 ctcccaccca acttttgcca attgcaacag tgacttctgt acaatttgca acagatacca
     2341 acgtgtcgag tcagcacaaa tggcgaggaa ggttcgcgtg cattgcttgc gcgcctcttt
     2401 gtcatgggcg cacgctgatt tgtgtgtgat ttgcaccaaa tttcctgccg acacaacttg
     2461 ggcccgtacc acacgacccg tttctggacc cgtttctggc ccagaaacgc gaatgttgcg
     2521 aaagatgcga ccacgtgctc gctgtcgaca acacaaactc ccgcacaatc cccatgaaaa
     2581 aaaaagtgct tgctgggctt gtgtctggcc tgtgtgcgtg gcgtggccgc gtgtggcttc
     2641 gttcttgctt ctgctcactc agcctgttac acgcagagtg gctgcgagaa acaaggagag
     2701 gggtttgcgg cattggcgca tgtgcatctg cctttttcat tgagtcagtc tgcagctcaa
     2761 gaggttggtt tcgtctacgg atatgagtga gcttctatgg atgatgagag gtggtggtgg
     2821 ttgtatggtt ggtggttggt ggttggtgca aggcgagcca cgcgaagagc gacaaggatg
     2881 agggagagga gaggagacta ggaaagaggt gaagggaagg gaagggcgag caatgacgac
     2941 gagataaaag acagccacca ccgcacccgc acaaaagcat tgctcgcacg catggtcgtg
     3001 tgttatgttt cacaccacca acataaccaa tacaagccat caccaataca agccagcgct
     3061 caagcacaca cgcacgcacg cacacacaat acacacgcac gtgaaacgtt cacattgtcc
     3121 gctcacaaat gcctttccct tctcgtcgct ttccttgcct ttttttcttc ccctcctgac
     3181 ttccttcctg ccttccttcc ttcctcccct tctccgtcca ccaacaccat ttatttctct
     3241 gctctttttc ctcgtgccac gtgagcagtc ttttcgcgtc gtcttcttct ttgaccccgt
     3301 ttaccgtcaa caccatcaac aaccagcaac atgagtggcg acataggctt tgccaacctc
     3361 cccaatcaaa tccatcgtga ggtcacgctc aaggggtttg acttcacttt gatggtcgtt
     3421 gggcggtctg gattgggaaa atcgacgttt gtcaacaccc tgttctccac cgacatctat
     3481 gaagaggaca agccctatcc cccaccttgt gctcgcattc ccagcacaac ctctgtgcag
     3541 gagagcgtga caacgctcat tgagaacgac gttcagctca cgctcacgct cgtggacacg
     3601 ccgggctttg gaaacggcat caacaacaca gactgctggg tccccattga ggagtatgtt
     3661 gacaatgcgt tctccaagta cgagggcgag gagtcgaaga tcagccgcac ggatatccgt
     3721 gattgccgcg tccactgctg cctctacttc ctcactcctt gcatccacgg cgtccgtccc
     3781 ctcgacattg aagtcatgaa gcgcctgcac acgaaggtga acatcatccc cgtcattgca
     3841 aaggcggatt cgctcacagt ggacgagtgc cagctcctca aggaacgcgt gcgcgccacg
     3901 ctggcggagc acaacatcaa cgtgtttgag tttcccgaag atgacgatga tgatgaagag
     3961 gtgcaagacg acgttgcaaa cgccaggtct gcgtggccgt ttgctgtatc ggcctcaaca
     4021 caggtactct tcgcagatga taaggaaatg cgtggccgcg cctacccctg gggttacgtc
     4081 gaagtcgaca atccgcagca ctcggacttc gttggcctgc gcaacctaat catcagaacg
     4141 tacatgcagc agctgattga cacgacgaac aacgtccact acgagcgttt ccgtcagcag
     4201 cgcctctcca gcctctctct tgacacatcc atcagtggaa gccccaaatc gtctggcggt
     4261 agcggtggct ccgtctccaa gggcgaggag accacgatgg gcgtcatcaa gcccgacatg
     4321 aagatcaagc tcaagatgga gggcaacgtc aacggccacg ccttcgtcat cgagggcgag
     4381 ggcgagggca agccctacga cggcaccaac accatcaacc tcgaggtcaa ggagggcgcc
     4441 cccctcccct tctcctacga catcctcacc accgccttcg cctacggcaa ccgcgccttc
     4501 accaagtacc ccgacgacat ccccaactac ttcaagcagt ccttccccga gggctactcc
     4561 tgggagcgca ccatgacctt cgaggacaag ggcatcgtca aggtcaagtc cgacatctct
     4621 atggaggagg actccttcat ctacgagatc cacctcaagg gcgagaactt cccccccaac
     4681 ggccccgtca tgcagaagaa gaccaccggc tgggacgcct ccaccgagcg catgtacgtc
     4741 cgcgacggcg tcctcaaggg cgacgtgaag cacaagctcc tcctcgaggg cggcggccac
     4801 caccgcgtcg acttcaagac catctaccgc gccaagaagg ccgtcaagct ccccgactac
     4861 cacttcgtcg accaccgcat cgagatcctc aaccacaaca aggactacaa caaggtcacc
     4921 gtctacgagt ccgccgtcgc ccgcaactcc accgacggca tggacgagct gtacaaggct
     4981 taagcgcctc gttggtcatt gccaccttgt tgcctctttt gctgttgtcc tcgtgtcgac
     5041 gcgtgccacg tgcgcttgtt gtgagttgat tctgcttctt gtgcacgtca tgccgagccc
     5101 ttgggccccg caataaaccc actatgccag tgtaactatc cttcccccca atgtataact
     5161 gtgacgatca atgagcggtg cccgtctaga ggatccccgg gtaccgagct cgaattcact
     5221 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     5281 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     5341 ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg atgcggtatt ttctccttac
     5401 gcatctgtgc ggtatttcac accgcatatg gtgcactctc agtacaatct gctctgatgc
     5461 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     5521 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     5581 gaggttttca ccgtcatcac cgaaacgcgc gagacga
//
