LOCUS       sequence-210507-        4308 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 4308)
  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 4308)
  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: NK612
FEATURES             Location/Qualifiers
     source          1..4308
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(12..30)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        98..202
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(203..271,272..1063)
                     /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(421..440)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1234..1822
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1723..1742
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1976..1993
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2110..2131
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2146..2151,2152..2169,2170..2176)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2184..2200
                     /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     2189..2211
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2208..2224
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2208..2224
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2899..2949
                     /codon_start=1
                     /product="peptide that binds filamentous actin (Riedl et 
                     al., 2008)"
                     /label=Lifeact
                     /note="first 17 amino acids of budding yeast Abp140"
                     /translation="MGVADLIKKFESISKEE"
     primer_bind     complement(3916..3933)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3916..3932)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3925..3947)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4141..4160)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4260..4282
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
       61 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
      121 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
      181 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
      241 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
      301 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
      361 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
      421 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
      481 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
      541 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
      601 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
      661 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
      721 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
      781 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
      841 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
      901 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
      961 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     1021 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     1081 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     1141 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     1201 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     1261 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     1321 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     1381 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     1441 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     1501 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     1561 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     1621 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     1681 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     1741 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     1801 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     1861 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     1921 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     1981 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     2041 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     2101 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct
     2161 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta
     2221 tgaccatgat tacgccaagc ttgtaacaaa ctgccctgca cctcgtctgc cacctcggtt
     2281 gccaccttgt cctcgttccc caccaccttg tcctcgtccg ctgccagcat cgcgatcagc
     2341 gtgcttgcgc ttggcgctgc tgctgccgga cttcccgtta tcggccatac tttttgtggt
     2401 gtttccaggc gagatgcgct gttggcaaag caaaatggca aaacggcctt tgccacacgc
     2461 catcctcgcc tcacctcgac gtgcggcttt cctaaacagg aaacctgacg aaacgaattg
     2521 agcgcattgg cacacgtgtg ctgtgcgctt tgtgtcgctg catgctatcc accacgcacc
     2581 acccaatcag tccaactgtg acgtgtcacg cctaggcgaa aaagcggcga gagcgccaaa
     2641 gcaaacgcca aacaaaaacc aaagaagagc aaacgcagcg aactgatgtg agacaagggg
     2701 gcggtgcaat gcgtcactga acgtgcaaaa gccaaaaaca accaagaagc agctgcacaa
     2761 cacaacaaca acaacaacaa caacaacacc ttgttgtcgc agctctacgt ttggcatttt
     2821 cctgttgttc tctcttgcga tcagtccgtg tctcttttct cccccttttc ccagaccaca
     2881 accacaaaac aaccagccat gggtgttgct gaccttatta agaaattcga gtctatctcc
     2941 aaggaagagg gcggtagcgg tggatctatg gtctccaagg gcgaggagga caacatggct
     3001 atcatcaagg agttcatgcg cttcaaggtc cacatggagg gctccgtcaa cggccacgag
     3061 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctc
     3121 aaggtgacga agggcggccc cctccccttc gcctgggaca tcctctcccc ccagttcatg
     3181 tacggctcca aggcttacgt caagcacccc gccgacatcc ccgactacct caagctctcc
     3241 ttccccgagg gcttcaagtg ggagcgcgtc atgaacttcg aggacggcgg cgtcgtcacc
     3301 gtcacccagg actcctctct tcaggacggc gagttcatct acaaggtcaa gctgcgcggc
     3361 accaacttcc cctccgacgg ccccgtcatg cagaagaaga cgatgggttg ggaggcttcc
     3421 tccgagcgca tgtaccccga ggacggcgcc ctcaagggcg agatcaagca gcgcctcaag
     3481 ctcaaggacg gcggccacta cgacgccgag gtcaagacca cctacaaggc caagaagccc
     3541 gtccagctcc ccggcgccta caacgtcaac atcaagctcg acatcacctc ccacaacgag
     3601 gactacacta tcgttgagca gtatgagcgt gccgagggtc gtcacagcac gggcggtatg
     3661 gacgagcttt acaagtctta agcgcctcgt tggtcattgc caccttgttg cctcttttgc
     3721 tgttgtcctc gtgtcgacgc gtgccacgtg cgcttgttgt gagttgattc tgcttcttgt
     3781 gcacgtcatg ccgagccctt gggccccgca ataaacccac tatgccagtg taactatcct
     3841 tccccccaat gtataactgt gacgatcaat gagcggtgcc cgtctagagg atccccgggt
     3901 accgagctcg aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     3961 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     4021 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat
     4081 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt gcactctcag
     4141 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     4201 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     4261 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgc
//
