LOCUS       sequence-210682-        4678 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 4678)
  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 4678)
  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..4678
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(15..33)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        101..205
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(206..274,275..1066)
                     /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(424..443)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1237..1825
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1726..1745
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1979..1996
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2113..2134
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2149..2154,2155..2172,2173..2179)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2187..2203
                     /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     2192..2214
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2211..2227
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2211..2227
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4289..4306)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4289..4305)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4298..4320)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4514..4533)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4633..4655
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
       61 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
      121 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
      181 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
      241 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
      301 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
      361 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
      421 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
      481 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
      541 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
      601 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
      661 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
      721 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
      781 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
      841 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
      901 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
      961 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     1021 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     1081 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     1141 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     1201 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     1261 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     1321 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     1381 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     1441 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     1501 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     1561 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     1621 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     1681 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     1741 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     1801 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     1861 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     1921 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     1981 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     2041 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     2101 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat
     2161 gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag
     2221 ctatgaccat gattacgcca agcttgtaac gactgcagtc ttgttgtcgt ttgtcgtcgt
     2281 cgttctctgt ctcccaccca acttttgcca attgcaacag tgacttctgt acaatttgca
     2341 acagatacca acgtgtcgag tcagcacaaa tggcgaggaa ggttcgcgtg cattgcttgc
     2401 gcgcctcttt gtcatgggcg cacgctgatt tgtgtgtgat ttgcaccaaa tttcctgccg
     2461 acacaacttg ggcccgtacc acacgacccg tttctggacc cgtttctggc ccagaaacgc
     2521 gaatgttgcg aaagatgcga ccacgtgctc gctgtcgaca acacaaactc ccgcacaatc
     2581 cccatgaaaa aaaaagtgct tgctgggctt gtgtctggcc tgtgtgcgtg gcgtggccgc
     2641 gtgtggcttc gttcttgctt ctgctcactc agcctgttac acgcagagtg gctgcgagaa
     2701 acaaggagag gggtttgcgg cattggcgca tgtgcatctg cctttttcat tgagtcagtc
     2761 tgcagctcaa gaggttggtt tcgtcgacgg atatgagtga gcttctatgg atgatgagag
     2821 gtggtggtgg ttgtatggtt ggtggttggt ggttggtgca aggcgagcca cgcgaagagc
     2881 gacaaggatg agggagagga gaggagacta ggaaagaggt gaagggaagg gaagggcgag
     2941 caatgacgac gagataaaag acagccacca ccgcacccgc acaaaagcat tgctcgcacg
     3001 catggtcgtg tgttatgttt cacaccacca acataaccaa tacaagccat caccaataca
     3061 agccagcgct caagcacaca cgcacgcacg cacacacaat acacacgcac gtgaaacgtt
     3121 cacattgtcc gctcacaaat gcctttccct tctcgtcgct ttccttgcct ttttttcttc
     3181 ccctcctgac ttccttcctg ccttccttcc ttcctcccct tctccgtcca ccaacaccat
     3241 ttatttctct gctctttttc ctcgtgccac gtgagcagtc ttttcgcgtc gtcttcttct
     3301 ttgaccccgt ttaccgtcaa caccatcaac aaccagcaac atggtctcca agggcgagga
     3361 gaccacgatg ggcgtcatca agcccgacat gaagatcaag ctcaagatgg agggcaacgt
     3421 caacggccac gccttcgtca tcgagggcga gggcgagggc aagccctacg acggcaccaa
     3481 caccatcaac ctcgaggtca aggagggcgc ccccctcccc ttctcctacg acatcctcac
     3541 caccgccttc gcctacggca accgcgcctt caccaagtac cccgacgaca tccccaacta
     3601 cttcaagcag tccttccccg agggctactc ctgggagcgc accatgacct tcgaggacaa
     3661 gggcatcgtc aaggtcaagt ccgacatctc tatggaggag gactccttca tctacgagat
     3721 ccacctcaag ggcgagaact tcccccccaa cggccccgtc atgcagaaga agaccaccgg
     3781 ctgggacgcc tccaccgagc gcatgtacgt ccgcgacggc gtcctcaagg gcgacgtgaa
     3841 gcacaagctc ctcctcgagg gcggcggcca ccaccgcgtc gacttcaaga ccatctaccg
     3901 cgccaagaag gccgtcaagc tccccgacta ccacttcgtc gaccaccgca tcgagatcct
     3961 caaccacaac aaggactaca acaaggtcac cgtctacgag tccgccgtcg cccgcaactc
     4021 caccgacggc atggacgagc tgtacaaggc ttaagcgcct cgttggtcat tgccaccttg
     4081 ttgcctcttt tgctgttgtc ctcgtgtcga cgcgtgccac gtgcgcttgt tgtgagttga
     4141 ttctgcttct tgtgcacgtc atgccgagcc cttgggcccc gcaataaacc cactatgcca
     4201 gtgtaactat ccttcccccc aatgtataac tgtgacgatc aatgagcggt gcccgtctag
     4261 aggatccccg ggtaccgagc tcgaattcac tggccgtcgt tttacaacgt cgtgactggg
     4321 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     4381 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     4441 aatggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat
     4501 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
     4561 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     4621 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacg
//
