LOCUS       sequence-194139-        3740 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses S. meditearranea GRSF1-4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3740)
  AUTHORS   Iyer H, Issigonis M, Sharma PP, Extavour CG, Newmark PA
  TITLE     A premeiotic function for boule in the planarian Schmidtea 
            mediterranea.
  JOURNAL   Proc Natl Acad Sci U S A. 2016 Jun 21;113(25):E3509-18. doi: 
            10.1073/pnas.1521341113. Epub 2016 Jun 2.
  PUBMED    27330085
REFERENCE   2  (bases 1 to 3740)
  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: 
            "10.1073/pnas.1521341113"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2016-06-21- 21"; volume: "113"; issue: "25"; pages: 
            "E3509-18"
COMMENT     Sequence Label: GRSF1-4
FEATURES             Location/Qualifiers
     source          1..3740
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(1..48)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        62..80
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        84..102
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        complement(619..637)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(620..637)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        complement(641..659)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      673..720
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(732..749)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(903..1491)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(983..1002)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1662..2453,2454..2522))
                     /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     2285..2304
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(2540..3334)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2651..2670)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3261..3280
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt tatgctaggc gcgccggatc
       61 ctaatacgac tcactatagg gagaattaac cctcactaaa gggagacctt gagaaaggcc
      121 acgaagagaa agatcaagaa gccaagttca tgagcgaatt aacgaaaatt atgtattgag
      181 gcttaaagga ttaccatata atgcatcaaa gacagatatt aaagaattct ttcaaggact
      241 gaaaattttg ccggatggaa tgagtattct cgttgatcat aatggaaggg cgctaggaga
      301 tgcttttgtt caatttgcaa cgtaccatga tgcaaaactt gccaaggaac ggaatggaaa
      361 taaaattgct cacaggtaca ttgaagtatt tgatagtaat attgaagaag ccaataatga
      421 aattcgtcga caaatggagt ttaatgaaat gcatgtgaaa aggtttgaaa aatctccgcc
      481 accatcaaga ggacctaggt ccatggatga cggaatacct tatcctgatt cacagagatt
      541 ttacggtaga attccagtaa ttgtcaatat gaaaggagtt tcaataagta cacgcccaga
      601 tgatgtttta aggtcacttc tatagtgtca cctaaatctc cctatagtga gtcgtattag
      661 gatccggcgc gcctagcata accccttggg gcctctaaac gggtcttgag gggttttttg
      721 acgcgtgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
      781 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
      841 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
      901 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
      961 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     1021 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     1081 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     1141 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     1201 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     1261 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     1321 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     1381 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     1441 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     1501 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     1561 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     1621 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     1681 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg gctccccgtc
     1741 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     1801 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     1861 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     1921 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     1981 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     2041 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     2101 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     2161 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     2221 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     2281 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     2341 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     2401 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     2461 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     2521 atactcttcc tttttcaatt cagaagaact cgtcaagaag gcgatagaag gcgatgcgct
     2581 gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa
     2641 gctcttcagc aatatcacgg gtagccaacg ctatgtcctg atagcggtcc gccacaccca
     2701 gccggccaca gtcgatgaat ccagaaaagc ggccattttc caccatgata ttcggcaagc
     2761 aggcatcgcc atgggtcacg acgagatcct cgccgtcggg catgcgcgcc ttgagcctgg
     2821 cgaacagttc ggctggcgcg agcccctgat gctcttcgtc cagatcatcc tgatcgacaa
     2881 gaccggcttc catccgagta cgtgctcgct cgatgcgatg tttcgcttgg tggtcgaatg
     2941 ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc atcagccatg atggatactt
     3001 tctcggcagg agcaaggtgg gatgacagga gatcctgccc cggcacttcg cccaatagca
     3061 gccagtccct tcccgcttca gtgacaacgt cgagcacagc tgcgcaagga acgcccgtcg
     3121 tggccagcca cgatagccgc gctgcctcgt cctgcagttc attcagggca ccggacaggt
     3181 cggtcttgac aaaaagaacc gggcgcccct gcgctgacag ccggaacacg gcggcatcag
     3241 agcagccgat tgtctgttgt gcccagtcat agccgaatag cctctccacc caagcggccg
     3301 gagaacctgc gtgcaatcca tcttgttcaa tcatgcgaaa cgatcctcat cctgtctctt
     3361 gatcagatct tgatcccctg cgccatcaga tcattggcgg caagaaagcc atccagttta
     3421 ctttgcaggg cttcccaacc ttaccagagg gcgccccagc tggcaattcc ggttcgcttg
     3481 ctgtccataa aaccgcccag tctagctatc gccatgtaag cccactgcaa gctacctgct
     3541 ttctctttgc gcttgcgttt tcccttgtcc agatagccca gtagctgaca ttcatccggg
     3601 gtcagcaccg tttctgcgga ctggctttct acgtgttccg cttcctttag cagcccttgc
     3661 gccctgaatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     3721 ataggccgaa atcgacgcgt
//
