LOCUS       sequence-210524-        5932 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 5932)
  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 5932)
  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: NK641
FEATURES             Location/Qualifiers
     source          1..5932
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(8..26)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        94..198
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(199..267,268..1059)
                     /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(417..436)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1230..1818
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1719..1738
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1972..1989
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2106..2127
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2142..2147,2148..2165,2166..2172)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2180..2196
                     /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     2185..2207
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2204..2220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2204..2220
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5536..5553)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5536..5552)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5545..5567)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5761..5780)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5880..5902
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
       61 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
      121 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
      181 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
      241 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
      301 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
      361 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
      421 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
      481 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
      541 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
      601 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
      661 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
      721 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
      781 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
      841 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
      901 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
      961 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     1021 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     1081 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     1141 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     1201 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     1261 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     1321 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     1381 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     1441 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     1501 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     1561 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     1621 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     1681 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     1741 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     1801 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     1861 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     1921 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     1981 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     2041 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     2101 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
     2161 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
     2221 catgattacg ccaagcttgt aacgactgca gtcttgttgt cgtttgtcgt cgtcgttctc
     2281 tgtctcccac ccaacttttg ccaattgcaa cagtgacttc tgtacaattt gcaacagata
     2341 ccaacgtgtc gagtcagcac aaatggcgag gaaggttcgc gtgcattgct tgcgcgcctc
     2401 tttgtcatgg gcgcacgctg atttgtgtgt gatttgcacc aaatttcctg ccgacacaac
     2461 ttgggcccgt accacacgac ccgtttctgg acccgtttct ggcccagaaa cgcgaatgtt
     2521 gcgaaagatg cgaccacgtg ctcgctgtcg acaacacaaa ctcccgcaca atccccatga
     2581 aaaaaaaagt gcttgctggg cttgtgtctg gcctgtgtgc gtggcgtggc cgcgtgtggc
     2641 ttcgttcttg cttctgctca ctcagcctgt tacacgcaga gtggctgcga gaaacaagga
     2701 gaggggtttg cggcattggc gcatgtgcat ctgccttttt cattgagtca gtctgcagct
     2761 caagaggttg gtttcgtcga cggatatgag tgagcttcta tggatgatga gaggtggtgg
     2821 tggttgtatg gttggtggtt ggtggttggt gcaaggcgag ccacgcgaag agcgacaagg
     2881 atgagggaga ggagaggaga ctaggaaaga ggtgaaggga agggaagggc gagcaatgac
     2941 gacgagataa aagacagcca ccaccgcacc cgcacaaaag cattgctcgc acgcatggtc
     3001 gtgtgttatg tttcacacca ccaacataac caatacaagc catcaccaat acaagccagc
     3061 gctcaagcac acacgcacgc acgcacacac aatacacacg cacgtgaaac gttcacattg
     3121 tccgctcaca aatgcctttc ccttctcgtc gctttccttg cctttttttc ttcccctcct
     3181 gacttccttc ctgccttcct tccttcctcc ccttctccgt ccaccaacac catttatttc
     3241 tctgctcttt ttcctcgtgc cacgtgagca gtcttttcgc gtcgtcttct tcttttaccc
     3301 cgtttaccgt caacaccatc aacaaccagc aacatgagtg gcgacatagg ctttgccaac
     3361 ctccccaatc aaatccatcg tgaggtcacg ctcaaggggt ttgacttcac tttgatggtc
     3421 gttgggcggt ctggattggg aaaatcgacg tttgtcaaca ccctgttctc caccgacatc
     3481 tatgaagagg acaagcccta tcccccacct tgtgctcgca ttcccagcac aacctctgtg
     3541 caggagagcg tgacaacgct cattgagaac gacgttcagc tcacgctcac gctcgtggac
     3601 acgccgggct ttggaaacgg catcaacaac acagactgct gggtccccat tgaggagtat
     3661 gttgacaatg cgttctccaa gtacgagggc gaggagtcga agatcagccg cacggatatc
     3721 cgtgattgcc gcgtccactg ctgcctctac ttcctcactc cttgcatcca cggcgtccgt
     3781 cccctcgaca ttgaagtcat gaagcgcctg cacacgaagg tgaacatcat ccccgtcatt
     3841 gcaaaggcgg attcgctcac agtggacgag tgccagctcc tcaaggaacg cgtgcgcgcc
     3901 acgctggcgg agcacaacat caacgtgttt gagtttcccg aagatgacga tgatgatgaa
     3961 gaggtgcaag acgacgttgc aaacgccagg tctgcgtggc cgtttgctgt atcggcctca
     4021 acacaggtac tcttcgcaga tgataaggaa atgcgtggcc gcgcctaccc ctggggttac
     4081 gtcgaagtcg acaatccgca gcactcggac ttcgttggcc tgcgcaacct aatcatcaga
     4141 acgtacatgc agcagctgat tgacacgacg aacaacgtcc actacgagcg tttccgtcag
     4201 cagcgcctct ccagcctctc tcttgacaca tccatcagtg gaagccccaa atcggatccc
     4261 aacagccaaa acttcatcga acagctcgaa ggacagctca aacaacacga gaagaagctt
     4321 gtgaagatgg agcgagagat gacagaagtc tttgagaagc gcgtgcgcga aaaacagcag
     4381 aaaatcaaga cgcaagaggc gcagctgctg aaggagcacg aggagtttga agaaaagttg
     4441 aagaagcagc aaagggagct tgagatgcag cgcgagcagt ttgagcggga gaagaaggag
     4501 tacgaggccg aggaggagaa gcgaaaggcc gcacaggcgc agtccactgg ccggcgcaag
     4561 aagttcttct ctggcggtag cggtggctcc gtctccaagg gcgaggagac cacgatgggc
     4621 gtcatcaagc ccgacatgaa gatcaagctc aagatggagg gcaacgtcaa cggccacgcc
     4681 ttcgtcatcg agggcgaggg cgagggcaag ccctacgacg gcaccaacac catcaacctc
     4741 gaggtcaagg agggcgcccc cctccccttc tcctacgaca tcctcaccac cgccttcgcc
     4801 tacggcaacc gcgccttcac caagtacccc gacgacatcc ccaactactt caagcagtcc
     4861 ttccccgagg gctactcctg ggagcgcacc atgaccttcg aggacaaggg catcgtcaag
     4921 gtcaagtccg acatctctat ggaggaggac tccttcatct acgagatcca cctcaagggc
     4981 gagaacttcc cccccaacgg ccccgtcatg cagaagaaga ccaccggctg ggacgcctcc
     5041 accgagcgca tgtacgtccg cgacggcgtc ctcaagggcg acgtgaagca caagctcctc
     5101 ctcgagggcg gcggccacca ccgcgtcgac ttcaagacca tctaccgcgc caagaaggcc
     5161 gtcaagctcc ccgactacca cttcgtcgac caccgcatcg agatcctcaa ccacaacaag
     5221 gactacaaca aggtcaccgt ctacgagtcc gccgtcgccc gcaactccac cgacggcatg
     5281 gacgagctgt acaaggctta agcgcctcgt tggtcattgc caccttgttg cctcttttgc
     5341 tgttgtcctc gtgtcgacgc gtgccacgtg cgcttgttgt gagttgattc tgcttcttgt
     5401 gcacgtcatg ccgagccctt gggccccgca ataaacccac tatgccagtg taactatcct
     5461 tccccccaat gtataactgt gacgatcaat gagcggtgcc cgtctagagg atccccgggt
     5521 accgagctcg aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     5581 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     5641 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat
     5701 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt gcactctcag
     5761 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     5821 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     5881 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ga
//
