LOCUS       sequence-210669-        4275 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 4275)
  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 4275)
  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: NK644
FEATURES             Location/Qualifiers
     source          1..4275
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(6..24)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        92..196
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(197..265,266..1057)
                     /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(415..434)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1228..1816
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1717..1736
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1970..1987
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2104..2125
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2140..2145,2146..2163,2164..2170)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2178..2194
                     /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     2183..2205
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2202..2218
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2202..2218
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3877..3894)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3877..3893)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3886..3908)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4102..4121)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4221..4243
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
       61 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
      121 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
      181 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
      241 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
      301 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
      361 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
      421 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
      481 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
      541 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
      601 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
      661 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
      721 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
      781 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
      841 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
      901 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
      961 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     1021 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     1081 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     1141 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     1201 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     1261 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     1321 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     1381 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     1441 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     1501 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     1561 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     1621 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     1681 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     1741 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     1801 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     1861 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     1921 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     1981 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     2041 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     2101 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     2161 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
     2221 tgattacgcc aagcttgtaa caaactgccc tgcacctcgt ctgccacctc ggttgccacc
     2281 ttgtcctcgt tccccaccac cttgtcctcg tccgctgcca gcatcgcgat cagcgtgctt
     2341 gcgcttggcg ctgctgctgc cggacttccc gttatcggcc atactttttg tggtgtttcc
     2401 aggcgagatg cgctgttggc aaagcaaaat ggcaaaacgg cctttgccac acgccatcct
     2461 cgcctcacct cgacgtgcgg ctttcctaaa caggaaacct gacgaaacga attgagcgca
     2521 ttggcacacg tgtgctgtgc gctttgtgtc gctgcatgct atccaccacg caccacccaa
     2581 tcagtccaac tgtgacgtgt cacgcctagg cgaaaaagcg gcgagagcgc caaagcaaac
     2641 gccaaacaaa aaccaaagaa gagcaaacgc agcgaactga tgtgagacaa gggggcggtg
     2701 caatgcgtca ctgaacgtgc aaaagccaaa aacaaccaag aagcagctgc acaacacaac
     2761 aacaacaaca acaacaacaa caccttgttg tcgcagctct acgtttggca ttttcctgtt
     2821 gttctctctt gcgatcagtc cgtgtctctt ttctccccct tttcccagac cacaaccaca
     2881 aaacaaccag ccatggtctc caagggcgag gaggacaaca tggctatcat caaggagttc
     2941 atgcgcttca aggtccacat ggagggctcc gtcaacggcc acgagttcga gatcgagggc
     3001 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctcaaggt gacgaagggc
     3061 ggccccctcc ccttcgcctg ggacatcctc tccccccagt tcatgtacgg ctccaaggct
     3121 tacgtcaagc accccgccga catccccgac tacctcaagc tctccttccc cgagggcttc
     3181 aagtgggagc gcgtcatgaa cttcgaggac ggcggcgtcg tcaccgtcac ccaggactcc
     3241 tctcttcagg acggcgagtt catctacaag gtcaagctgc gcggcaccaa cttcccctcc
     3301 gacggccccg tcatgcagaa gaagacgatg ggttgggagg cttcctccga gcgcatgtac
     3361 cccgaggacg gcgccctcaa gggcgagatc aagcagcgcc tcaagctcaa ggacggcggc
     3421 cactacgacg ccgaggtcaa gaccacctac aaggccaaga agcccgtcca gctccccggc
     3481 gcctacaacg tcaacatcaa gctcgacatc acctcccaca acgaggacta caccatcgtc
     3541 gagcagtacg agcgtgccga gggccgccac agcacgggcg gtatggacga gctttacaag
     3601 tctaccgcaa agaccggtgg tgggtgctgc acgctgctct aagcgcctcg ttggtcattg
     3661 ccaccttgtt gcctcttttg ctgttgtcct cgtgtcgacg cgtgccacgt gcgcttgttg
     3721 tgagttgatt ctgcttcttg tgcacgtcat gccgagccct tgggccccgc aataaaccca
     3781 ctatgccagt gtaactatcc ttccccccaa tgtataactg tgacgatcaa tgagcggtgc
     3841 ccgtctagag gatccccggg taccgagctc gaattcactg gccgtcgttt tacaacgtcg
     3901 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
     3961 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     4021 gaatggcgaa tggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     4081 ccgcatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
     4141 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     4201 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     4261 gaaacgcgcg agacg
//
