LOCUS       sequence-210500-        6247 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 6247)
  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 6247)
  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: NK619
FEATURES             Location/Qualifiers
     source          1..6247
                     /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(5849..5866)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5849..5865)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5858..5880)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6074..6093)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6193..6215
                     /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 cgactgcagt cttgttgtcg tttgtcgtcg tcgttctctg
     2281 tctcccaccc aacttttgcc aattgcaaca gtgacttctg tacaatttgc aacagatacc
     2341 aacgtgtcga gtcagcacaa atggcgagga aggttcgcgt gcattgcttg cgcgcctctt
     2401 tgtcatgggc gcacgctgat ttgtgtgtga tttgcaccaa atttcctgcc gacacaactt
     2461 gggcccgtac cacacgaccc gtttctggac ccgtttctgg cccagaaacg cgaatgttgc
     2521 gaaagatgcg accacgtgct cgctgtcgac aacacaaact cccgcacaat ccccatgaaa
     2581 aaaaaagtgc ttgctgggct tgtgtctggc ctgtgtgcgt ggcgtggccg cgtgtggctt
     2641 cgttcttgct tctgctcact cagcctgtta cacgcagagt ggctgcgaga aacaaggaga
     2701 ggggtttgcg gcattggcgc atgtgcatct gcctttttca ttgagtcagt ctgcagctca
     2761 agaggttggt ttcgtcgacg gatatgagtg agcttctatg gatgatgaga ggtggtggtg
     2821 gttgtatggt tggtggttgg tggttggtgc aaggcgagcc acgcgaagag cgacaaggat
     2881 gagggagagg agaggagact aggaaagagg tgaagggaag ggaagggcga gcaatgacga
     2941 cgagataaaa gacagccacc accgcacccg cacaaaagca ttgctcgcac gcatggtcgt
     3001 gtgttatgtt tcacaccacc aacataacca atacaagcca tcaccaatac aagccagcgc
     3061 tcaagcacac acgcacgcac gcacacacaa tacacacgca cgtgaaacgt tcacattgtc
     3121 cgctcacaaa tgcctttccc ttctcgtcgc tttccttgcc tttttttctt cccctcctga
     3181 cttccttcct gccttccttc cttcctcccc ttctccgtcc accaacacca tttatttctc
     3241 tgctcttttt cctcgtgcca cgtgagcagt cttttcgcgt cgtcttcttc tttgaccccg
     3301 tttaccgtca acaccatcaa caaccagcaa catgtctgag ggcaagcagc acgtttccat
     3361 tgtcatctgc ggccacgtcg atgccggcaa gtcgaccacg accggccgtc tcatcttcga
     3421 gctcggcggc atccccgagc gtgagatgga gaagctcaag gccgaggccg acgctctcgg
     3481 caagggctcc ttcgcctttg ccttctacat ggaccgccag aaggaggagc gcgagcgtgg
     3541 tgtgaccatt gcctgcacca ccaaggagtt cttcacggag tccaagcact acaccgtcat
     3601 tgacgcccct ggccatcgtg acttcatcaa gaacatgatc accggtgcct cccaggccga
     3661 tgtcgccctc ctcatggttc cctccgacgg taacttcacc gccgccattg ccaagggcaa
     3721 ccacaaggcc ggtgtcgtcc agggccagac ccgccagcac gccgtcctca tcaaccttct
     3781 cggcgtcaag cagctcattg ttggcgtcaa caagatggac tgcgacgttg ccaagtactc
     3841 caaggagcgc tacgaggaga tccgcgacga gatgcgccac atgctcatca agtctggctg
     3901 gaagaaggcc tttgtcatgg agtctgtccc cgtgctcccc atctccggct ggatgggcga
     3961 caacctcatc accaagtcca agaacatgga ctggtggaag ggcgtcgacg tcaaggttga
     4021 gggcgagacc ctccacctcg agacgctcaa ggaggctctt gagaagatgg tccgcgtccc
     4081 ccagcgcgcc acggacaagc ccatgcgcac ccccatctcc ggtgtcttca agatcaaggg
     4141 tgtcggtgat gtcctcaccg gccgcgtcga gcagggtacc gtcaagcccg gcgacgaggt
     4201 tgtcttcctc cccacccaca ctgaggccaa cccctgcact ggcaagatct tcaccgtcga
     4261 gatgcaccac aagtctgtcg aggctgccgc cactggcgac aacgttggcc tcaacgtcaa
     4321 gggtctggac aagaagaaca tgccccgcgt cggtgatgtc atgatcctca agtctgacaa
     4381 ctccctcggc aagtgcaaga gcttccacgc ccaggtccag gtcatgaacc accctggtga
     4441 gctcaaggcc ggctactgcc ccatcgcctt tgtccgcaca tcccgctctg ccgtccgcat
     4501 ggccgagatt tgctggaaga ttggcaagga gaccggtggc tccaaggccg agaaccccgc
     4561 caacctcaag gccaacgagg tcgccgaggt caagttcgag ccccagcagc ctttcgtcgt
     4621 cgacaccttc aagtcctgcg agggtcttgg ccgcattgcc atcatggagg gtaacactgt
     4681 cgtcatgctc ggcaaggtca ccaagatgga gcgcgcttct ggcggtagcg gtggctccat
     4741 ggtctccaag ggcgaggagg acaacatggc tatcatcaag gagttcatgc gcttcaaggt
     4801 ccacatggag ggctccgtca acggccacga gttcgagatc gagggcgagg gcgagggccg
     4861 cccctacgag ggcacccaga ccgccaagct caaggtgacg aagggcggcc ccctcccctt
     4921 cgcctgggac atcctctccc cccagttcat gtacggctcc aaggcttacg tcaagcaccc
     4981 cgccgacatc cccgactacc tcaagctctc cttccccgag ggcttcaagt gggagcgcgt
     5041 catgaacttc gaggacggcg gcgtcgtcac cgtcacccag gactcctctc ttcaggacgg
     5101 cgagttcatc tacaaggtca agctgcgcgg caccaacttc ccctccgacg gccccgtcat
     5161 gcagaagaag acgatgggtt gggaggcttc ctccgagcgc atgtaccccg aggacggcgc
     5221 cctcaagggc gagatcaagc agcgcctcaa gctcaaggac ggcggccact acgacgccga
     5281 ggtcaagacc acctacaagg ccaagaagcc cgtccagctc cccggcgcct acaacgtcaa
     5341 catcaagctc gacatcacct cccacaacga ggactacacc atcgtcgagc agtacgagcg
     5401 tgccgagggc cgccacagca cgggcggtat ggacgagctt tacaagtctt aagagctccc
     5461 cccagcatta tcacgtctac tgtgtgtatg ttgtcgctct ctgtttgacc tctctcttgt
     5521 ctactatgtt gcctctctct ctttcttgat tgcacccctt tcctcgctac gactactacg
     5581 acttccgcta ccttgacttt gttggcttgt tgccaccttt tggtcttgcg tttcgtctgc
     5641 tcgtcactct tgtctctttt tgacgacttg tgtgtgatga cggcgtgagg ctgtgactga
     5701 ctgactctca acaacttcca ataaaaaccc ctacatcgca cgtgactgac tgcacattta
     5761 gtgaagggaa ggaaaagaag tgaggtgttt tcatgtacat gaggcttgtc gttgctctag
     5821 aggatccccg ggtaccgagc tcgaattcac tggccgtcgt tttacaacgt cgtgactggg
     5881 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     5941 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     6001 aatggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat
     6061 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
     6121 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     6181 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     6241 cgagacg
//
