LOCUS       sequence-210429-        5165 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 5165)
  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 5165)
  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: NK610
FEATURES             Location/Qualifiers
     source          1..5165
                     /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"
     primer_bind     complement(4773..4790)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4773..4789)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4782..4804)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4998..5017)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5117..5139
                     /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 ttgtaagtag tggtagtcgt cgaagacgtg gggtgcgggg
     2281 atgtgcttga cttggtggga gaggctggtg gaagtagctg atgaaaatga actccaccac
     2341 acgccaactg ccacttgaac cacggaaggc aggcagcagt gctgctgtgt gcgcttttcg
     2401 gcgcggcgcg cagccacaaa gcaacggcaa ttgtggcggg tgattcgttc gaacgctgct
     2461 tttcataatt ttcgccaatg tctgaatcaa tgtctttttc aagggtgcag aataacgaac
     2521 attatttttt attgtgtgga ttcagcgcca attgcgattt ggagatgcgc taagcgcaca
     2581 atcaccacca aacacgacat tccccgcttt cccgccaaac tgattctcgc gcgaaacctg
     2641 agatgtgcac cgcgccgtta caagtgcgac cacgcacacc tcttcttgtc atttatcatc
     2701 cgcatctcca agcaagcaag caagcaaacc aggaccccct ccctcgctcg tcacctcacc
     2761 acctcccaaa cttaaacttc acaaccagca caatggcccg cacgaagcag accgcccgca
     2821 agaccaccgg tggcaaggcc ccccgcaagc agctcgccac caaggctgcc cgcaagtccg
     2881 ccccctccac tggcggcgtg aagaagcccc atcgcttccg ccccggtacc gtcgctctgc
     2941 gtgagatccg ccgctaccag aagtccaccg agctgctgat ccgcaagctc cccttccagc
     3001 gcctcgtccg cgagattgcc caggacttca agaccgacct ccgcttccag tccaccgccg
     3061 tctctgccct gcaggaggcc gccgaggctt acctcgtcaa cctctttgag gacaccaacc
     3121 tctgcgccat ccacgcaaag cgcgtcacca tcatgcccaa ggacatccag ctggcccgcc
     3181 gcatccgtgg cgagcgctct ggcggtagcg gtggctccat ggtctccaag ggcgaggagg
     3241 acaacatggc tatcatcaag gagttcatgc gcttcaaggt ccacatggag ggctccgtca
     3301 acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga
     3361 ccgccaagct caaggtgacg aagggcggcc ccctcccctt cgcctgggac atcctctccc
     3421 cccagttcat gtacggctcc aaggcttacg tcaagcaccc cgccgacatc cccgactacc
     3481 tcaagctctc cttccccgag ggcttcaagt gggagcgcgt catgaacttc gaggacggcg
     3541 gcgtcgtcac cgtcacccag gactcctctc ttcaggacgg cgagttcatc tacaaggtca
     3601 agctgcgcgg caccaacttc ccctccgacg gccccgtcat gcagaagaag acgatgggtt
     3661 gggaggcttc ctccgagcgc atgtaccccg aggacggcgc cctcaagggc gagatcaagc
     3721 agcgcctcaa gctcaaggac ggcggccact acgacgccga ggtcaagacc acctacaagg
     3781 ccaagaagcc cgtccagctc cccggcgcct acaacgtcaa catcaagctc gacatcacct
     3841 cccacaacga ggactacacc atcgtcgagc agtacgagcg tgccgagggc cgccacagca
     3901 cgggcggtat ggacgagctt tacaagtctt aagcgtctca ccttctcaac atggctcgct
     3961 cgctcgatgt cgttctctga caccttgctt ggcttgtcgt catgaggttg ctttgtcatc
     4021 accaacgctg gttgcatttt gatctgtgac accttgaatc atcctccctc ccctcccttg
     4081 ttcttaatta cccacttggc ccttgctgtc tctcttggtc tgtgccacgg gttgtcgcct
     4141 ccgcgggctc aaattgtgct cacacgtctt ttatccatgc tcgtgttcct gagcacttga
     4201 cgctggcggt gtctttgtgc gctcgtgtgc atgggaacct ctctccctcc ctctccctgt
     4261 gtgtaatggc gttggcttac cgtggtatcg atttgcctct gcgtaatgac acgccgcaac
     4321 ggcgtttggg gagtttcccc ggcttgtcct gcctcgcaac ccttctcctt tggttcgttg
     4381 ttgcctctca atacaatgat caaaccgaac attaatgtga ggaccatgat ggcatcctgc
     4441 tttattcaag caagtggaat gaatgacaca gaaacagatt gatcgtcatt gtgatgccgt
     4501 gcgtgtattt gtggcggaga ggaacaaaag tcacaggtgg ctacatttac acaacttctt
     4561 gtcaatgcaa actcagtgag tcattgggta tcatgcgtgg tgaagaatga gggagtgctg
     4621 ccagcatctg aacgagcacc atcgtgccag ggcgcggctc ttggcaaact cgcgtgcgtc
     4681 catctcttcg tgatcatcgt catcatcgcg acgctgtgtc acatgtggga caatgaatgt
     4741 ctagaggatc cccgggtacc gagctcgaat tcactggccg tcgttttaca acgtcgtgac
     4801 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
     4861 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     4921 ggcgaatggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     4981 atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     5041 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     5101 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     5161 cgcgc
//
