LOCUS       sequence-210426-        4263 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 4263)
  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 4263)
  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: NK594
FEATURES             Location/Qualifiers
     source          1..4263
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(7..25)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        93..197
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(198..266,267..1058)
                     /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(416..435)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1229..1817
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1718..1737
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1971..1988
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2105..2126
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2141..2146,2147..2164,2165..2171)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2179..2195
                     /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     2184..2206
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2203..2219
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2203..2219
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3866..3883)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3866..3882)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3875..3897)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4091..4110)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4210..4232
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
       61 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
      121 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
      181 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
      241 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
      301 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
      361 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
      421 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
      481 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
      541 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
      601 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
      661 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
      721 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
      781 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
      841 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
      901 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
      961 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     1021 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     1081 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     1141 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     1201 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     1261 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     1321 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     1381 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     1441 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     1501 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     1561 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     1621 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     1681 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     1741 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     1801 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     1861 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     1921 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     1981 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     2041 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     2101 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg
     2161 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc
     2221 atgattacgc caagcttgta acaaactgcc ctgcacctcg tctgccacct cggttgccac
     2281 cttgtcctcg ttccccacca ccttgtcctc gtccgctgcc agcatcgcga tcagcgtgct
     2341 tgcgcttggc gctgctgctg ccggacttcc cgttatcggc catacttttt gtggtgtttc
     2401 caggcgagat gcgctgttgg caaagcaaaa tggcaaaacg gcctttgcca cacgccatcc
     2461 tcgcctcacc tcgacgtgcg gctttcctaa acaggaaacc tgacgaaacg aattgagcgc
     2521 attggcacac gtgtgctgtg cgctttgtgt cgctgcatgc tatccaccac gcaccaccca
     2581 atcagtccaa ctgtgacgtg tcacgcctag gcgaaaaagc ggcgagagcg ccaaagcaaa
     2641 cgccaaacaa aaaccaaaga agagcaaacg cagcgaactg atgtgagaca agggggcggt
     2701 gcaatgcgtc actgaacgtg caaaagccaa aaacaaccaa gaagcagctg cacaacacaa
     2761 caacaacaac aacaacaaca acaccttgtt gtcgcagctc tacgtttggc attttcctgt
     2821 tgttctctct tgcgatcagt ccgtgtctct tttctccccc ttttcccaga ccacaaccac
     2881 aaaacaacca gccatggtct ccaagggcga ggagctgatc aaggagaaca tgcacatgaa
     2941 gctctacatg gagggcaccg tcaacaacca ccacttcaag tgcacctccg agggcgaggg
     3001 caagccctac gagggcaccc agaccatgcg catcaaggtc gtcgagggcg gccccctccc
     3061 cttcgccttc gacatcctcg ccacctcctt catgtacggc tcccgcacct tcatcaacca
     3121 cacccagggc atccccgact tcttcaagca gtccttcccc gagggcttca cctgggagcg
     3181 cgtcaccacc tacgaggacg gcggcgtcct caccgccacc caggacacct ccctgcagga
     3241 cggctgcctc atctacaacg tcaagatccg cggcgtcaac ttcccctcca acggccccgt
     3301 catgcagaag aagaccctcg gctgggaggc caacaccgag atgctctacc ccgccgacgg
     3361 cggcctcgag ggccgcaccg acatggccct caagctcgtc ggcggcggcc acctcatctg
     3421 caacttcaag accacctacc gctccaagaa gcccgccaag aacctcaaga tgcccggcgt
     3481 ctactacgtc gaccaccgcc tcgagcgcat caaggaggcc gacaaggaga cctacgtcga
     3541 gcagcacgag gtcgccgtcg cccgctactg cgacctcccc tccaagctcg gccacaagct
     3601 caacggcatg gacgagctgt acaaggctta agcgcctcgt tggtcattgc caccttgttg
     3661 cctcttttgc tgttgtcctc gtgtcgacgc gtgccacgtg cgcttgttgt gagttgattc
     3721 tgcttcttgt gcacgtcatg ccgagccctt gggccccgca ataaacccac tatgccagtg
     3781 taactatcct tccccccaat gtataactgt gacgatcaat gagcggtgcc cgtctagagg
     3841 atccccgggt accgagctcg aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa
     3901 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
     3961 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     4021 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt
     4081 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
     4141 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     4201 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     4261 gac
//
