LOCUS       sequence-212162-        4305 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 4305)
  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 4305)
  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: NK613
FEATURES             Location/Qualifiers
     source          1..4305
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(9..27)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        95..199
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(200..268,269..1060)
                     /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(418..437)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1231..1819
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1720..1739
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1973..1990
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2107..2128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2143..2148,2149..2166,2167..2173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2181..2197
                     /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     2186..2208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2205..2221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2205..2221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3619..3639
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3646..3666
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(3910..3927)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3910..3926)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3919..3941)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4135..4154)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4254..4276
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
       61 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
      121 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
      181 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
      241 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
      301 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
      361 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
      421 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
      481 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
      541 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
      601 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
      661 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
      721 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
      781 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
      841 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
      901 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
      961 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     1021 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     1081 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     1141 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     1201 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     1261 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     1321 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     1381 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     1441 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     1501 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     1561 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     1621 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     1681 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     1741 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     1801 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     1861 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     1921 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     1981 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     2041 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     2101 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     2161 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
     2221 ccatgattac gccaagcttg taacaaactg ccctgcacct cgtctgccac ctcggttgcc
     2281 accttgtcct cgttccccac caccttgtcc tcgtccgctg ccagcatcgc gatcagcgtg
     2341 cttgcgcttg gcgctgctgc tgccggactt cccgttatcg gccatacttt ttgtggtgtt
     2401 tccaggcgag atgcgctgtt ggcaaagcaa aatggcaaaa cggcctttgc cacacgccat
     2461 cctcgcctca cctcgacgtg cggctttcct aaacaggaaa cctgacgaaa cgaattgagc
     2521 gcattggcac acgtgtgctg tgcgctttgt gtcgctgcat gctatccacc acgcaccacc
     2581 caatcagtcc aactgtgacg tgtcacgcct aggcgaaaaa gcggcgagag cgccaaagca
     2641 aacgccaaac aaaaaccaaa gaagagcaaa cgcagcgaac tgatgtgaga caagggggcg
     2701 gtgcaatgcg tcactgaacg tgcaaaagcc aaaaacaacc aagaagcagc tgcacaacac
     2761 aacaacaaca acaacaacaa caacaccttg ttgtcgcagc tctacgtttg gcattttcct
     2821 gttgttctct cttgcgatca gtccgtgtct cttttctccc ccttttccca gaccacaacc
     2881 acaaaacaac cagccatggt ctccaagggc gaggaggaca acatggctat catcaaggag
     2941 ttcatgcgct tcaaggtcca catggagggc tccgtcaacg gccacgagtt cgagatcgag
     3001 ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctcaa ggtgacgaag
     3061 ggcggccccc tccccttcgc ctgggacatc ctctcccccc agttcatgta cggctccaag
     3121 gcttacgtca agcaccccgc cgacatcccc gactacctca agctctcctt ccccgagggc
     3181 ttcaagtggg agcgcgtcat gaacttcgag gacggcggcg tcgtcaccgt cacccaggac
     3241 tcctctcttc aggacggcga gttcatctac aaggtcaagc tgcgcggcac caacttcccc
     3301 tccgacggcc ccgtcatgca gaagaagacg atgggttggg aggcttcctc cgagcgcatg
     3361 taccccgagg acggcgccct caagggcgag atcaagcagc gcctcaagct caaggacggc
     3421 ggccactacg acgccgaggt caagaccacc tacaaggcca agaagcccgt ccagctcccc
     3481 ggcgcctaca acgtcaacat caagctcgac atcacctccc acaacgagga ctacaccatc
     3541 gtcgagcagt acgagcgtgc cgagggccgc cacagcacgg gcggtatgga cgagctttac
     3601 aagggtggcg acggcagccc caagaaaaag cgcaaggtcg aggaccctaa aaagaaacgc
     3661 aaggtcgact cttaagcgcc tcgttggtca ttgccacctt gttgcctctt ttgctgttgt
     3721 cctcgtgtcg acgcgtgcca cgtgcgcttg ttgtgagttg attctgcttc ttgtgcacgt
     3781 catgccgagc ccttgggccc cgcaataaac ccactatgcc agtgtaacta tccttccccc
     3841 caatgtataa ctgtgacgat caatgagcgg tgcccgtcta gaggatcccc gggtaccgag
     3901 ctcgaattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
     3961 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
     4021 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgcc tgatgcggta
     4081 ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc tcagtacaat
     4141 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     4201 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     4261 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgag
//
