LOCUS       sequence-461122-        6833 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses RGECO1 (fluorescent calcium indicator) and pac (puromycin 
            resistance) in the Choanoflagellate, Salpingoeca rosetta.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6833)
  AUTHORS   Colgren J, Burkhardt P
  TITLE     Electrical signaling and coordinated behavior in the closest 
            relative of animals.
  JOURNAL   Sci Adv. 2025 Jan 10;11(2):eadr7434. doi: 10.1126/sciadv.adr7434. 
            Epub 2025 Jan 8.
  PUBMED    39772683
REFERENCE   2  (bases 1 to 6833)
  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"; journalName: "Sci Adv."; 
            date: "2025-01-10"; volume: "11(2):eadr7434. doi"; pages: " 
            10.1126/sciadv.adr7434. Epub 2025 Jan 8"
COMMENT     Sequence Label: pSrRGECO1
FEATURES             Location/Qualifiers
     source          1..6833
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        96..200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(201..269,270..1061)
                     /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(419..438)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1232..1820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1721..1740
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1974..1991
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2108..2129
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2144..2149,2150..2167,2168..2174)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2182..2198
                     /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     2187..2209
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2206..2222
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2206..2222
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4793..4813)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(6439..6456)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6439..6455)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6448..6470)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6664..6683)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6783..6805
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2101 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     2161 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     2221 accatgatta cgccaagctt gtaacaaact gccctgcacc tcgtctgcca cctcggttgc
     2281 caccttgtcc tcgttcccca ccaccttgtc ctcgtccgct gccagcatcg cgatcagcgt
     2341 gcttgcgctt ggcgctgctg ctgccggact tcccgttatc ggccatactt tttgtggtgt
     2401 ttccaggcga gatgcgctgt tggcaaagca aaatggcaaa acggcctttg ccacacgcca
     2461 tcctcgcctc acctcgacgt gcggctttcc taaacaggaa acctgacgaa acgaattgag
     2521 cgcattggca cacgtgtgct gtgcgctttg tgtcgctgca tgctatccac cacgcaccac
     2581 ccaatcagtc caactgtgac gtgtcacgcc taggcgaaaa agcggcgaga gcgccaaagc
     2641 aaacgccaaa caaaaaccaa agaagagcaa acgcagcgaa ctgatgtgag acaagggggc
     2701 ggtgcaatgc gtcactgaac gtgcaaaagc caaaaacaac caagaagcag ctgcacaaca
     2761 caacaacaac aacaacaaca acaacacctt gttgtcgcag ctctacgttt ggcattttcc
     2821 tgttgttctc tcttgcgatc agtccgtgtc tcttttctcc cccttttccc agaccacaac
     2881 cacaaaacaa ccagccatgg tggacagcag ccgccgcaag tggaacaagg ccggccacgc
     2941 cgtgcgcgcc atcggccgcc tgagcagccc ggtggtgagc gagcgcatgt acccggagga
     3001 cggcgccctg aagagcgaga tcaagaaggg cctgcgcctg aaggacggcg gccactacgc
     3061 cgccgaggtg aagacgacgt acaaggccaa gaagccggtg cagctgccgg gcgcctacat
     3121 cgtggacatc aagctggaca tcgtgagcca caacgaggac tacacgatcg tggagcagtg
     3181 cgagcgcgcc gagggccgcc acagcacggg cggcatggac gagctgtaca agggcggcac
     3241 gggcggcagc ctggtgagca agggcgagga ggacaacatg gccatcatca aggagttcat
     3301 gcgcttcaag gtgcacatgg agggcagcgt gaacggccac gagttcgaga tcgagggcga
     3361 gggcgagggc cgcccgtacg aggccttcca gacggccaag ctgaaggtga cgaagggcgg
     3421 cccgctgccg ttcgcctggg acatcctgag cccgcagttc atgtacggca gcaaggccta
     3481 catcaagcac ccggccgaca tcccggacta cttcaagctg agcttcccgg agggcttccg
     3541 ctgggagcgc gtgatgaact tcgaggacgg cggcatcatc cacgtgaacc aggacagcag
     3601 cctgcaggac ggcgtgttca tctacaaggt gaagctgcgc ggcacgaact tcccgccgga
     3661 cggcccggtg atgcagaaga agacgatggg ctgggaggcc acgcgcgacc agctgacgga
     3721 ggagcagatc gccgagttca aggaggcctt cagcctgttc gacaaggacg gcgacggcac
     3781 gatcacgacg aaggagctgg gcacggtgat gcgcagcctg ggccagaacc cgacggaggc
     3841 cgagctgcag gacatgatca acgaggtgga cgccgacggc gacggcacgt tcgacttccc
     3901 ggagttcctg acgatgatgg cccgcaagat gaacgacacg gacagcgagg aggagatccg
     3961 cgaggccttc cgcgtgttcg acaaggacgg caacggctac atcggcgccg ccgagctgcg
     4021 ccacgtgatg acggacctgg gcgagaagct gacggacgag gaggtggacg agatgatccg
     4081 cgtggccgac atcgacggcg acggccaggt gaactacgag gagttcgtgc agatgatgac
     4141 ggccaagtaa gagctccccc cagcattatc acgtctactg tgtgtatgtt gtcgctctct
     4201 gtttgacctc tctcttgtct actatgttgc ctctctctct ttcttgattg cacccctttc
     4261 ctcgctacga ctactacgac ttccgctacc ttgactttgt tggcttgttg ccaccttttg
     4321 gtcttgcgtt tcgtctgctc gtcactcttg tctctttttg acgacttgtg tgtgatgacg
     4381 gcgtgaggct gtgactgact gactctcaac aacttccaat aaaaacccct acatcgcacg
     4441 tgactgactg cacatttagt gaagggaagg aaaagaagtg aggtgttttc atgtacatga
     4501 ggcttgtcgt tgctcgggca ccgctcattg atcgtcacag ttatacattg gggggaagga
     4561 tagttacact ggcatagtgg gtttattgcg gggcccaagg gctcggcatg acgtgcacaa
     4621 gaagcagaat caactcacaa caagcgcacg tggcacgcgt cgacacgagg acaacagcaa
     4681 aagaggcaac aaggtggcaa tgaccaacga ggcgcaccgg gcttgcgggt catgcaccag
     4741 gtacgaggac cctcggggac ctccacgtcg gcggtgacgg tgaagccgag acgctcgtag
     4801 aaggggaggt tgcgaggagc ggaggtctcg aggaaagcag ggacaccagc acgctcagcg
     4861 gcctcgacgc cagggaggac gacggcagag ccgagaccct tgccctggtg gtcaggagag
     4921 acgccgacgg tggcgaggaa ccaggcgggc tccttggggc ggtggggggc gaggaggccc
     4981 tccatctgct gctgagcggc gagacgggag ccagacagct cagccatgcg ggggccgatc
     5041 tcagcgaaga cagcaccggc ctcgacagac tcaggggtgg tccagacagc gacagcagca
     5101 ccgtcgtcgg cgacccagac cttcccgatg tcgaggccga cgcgggtgag gaacagctcc
     5161 tgcagctcgg tgacacgctc gatgtggcgg tcagggtcga cggtgtgacg ggtagcaggg
     5221 tagtcagcga aggcagcggc gagggtacgg acagcacggg gcacgtcgtc acgggtagcg
     5281 aggcggacgg taggcttgta ctccgtacca gacatgttgc tggttgttga tggtgttgac
     5341 ggtaaacggg gtcaaagaag aagacgacgc gaaaagactg ctcacgtggc acgaggaaaa
     5401 agagcagaga aataaatggt gttggtggac ggagaagggg aggaaggaag gaaggcagga
     5461 aggaagtcag gaggggaaga aaaaaaggca aggaaagcga cgagaaggga aaggcatttg
     5521 tgagcggaca atgtgaacgt ttcacgtgcg tgtgtattgt gtgtgcgtgc gtgcgtgtgt
     5581 gcttgagcgc tggcttgtat tggtgatggc ttgtattggt tatgttggtg gtgtgaaaca
     5641 taacacacga ccatgcgtgc gagcaatgct tttgtgcggg tgcggtggtg gctgtctttt
     5701 atctcgtcgt cattgctcgc ccttcccttc ccttcacctc tttcctagtc tcctctcctc
     5761 tccctcatcc ttgtcgctct tcgcgtggct cgccttgcac caaccaccaa ccaccaacca
     5821 tacaaccacc accacctctc atcatccata gaagctcact catatccgtc gacgaaacca
     5881 acctcttgag ctgcagactg actcaatgaa aaaggcagat gcacatgcgc caatgccgca
     5941 aacccctctc cttgtttctc gcagccactc tgcgtgtaac aggctgagtg agcagaagca
     6001 agaacgaagc cacacgcggc cacgccacgc acacaggcca gacacaagcc cagcaagcac
     6061 tttttttttc atggggattg tgcgggagtt tgtgttgtcg acagcgagca cgtggtcgca
     6121 tctttcgcaa cattcgcgtt tctgggccag aaacgggtcc agaaacgggt cgtgtggtac
     6181 gggcccaagt tgtgtcggca ggaaatttgg tgcaaatcac acacaaatca gcgtgcgccc
     6241 atgacaaaga ggcgcgcaag caatgcacgc gaaccttcct cgccatttgt gctgactcga
     6301 cacgttggta tctgttgcaa attgtacaga agtcactgtt gcaattggca aaagttgggt
     6361 gggagacaga gaacgacgac gacaaacgac aacaagactg cagtcgttac aggatccccg
     6421 ggtaccgagc tcgaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     6481 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     6541 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct
     6601 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct
     6661 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     6721 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     6781 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cga
//
