LOCUS       sequence-309584-        4318 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MosSCI SapTrap assembly: Cassette 4 for 4- or 5-cassette system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4318)
  AUTHORS   Fan X, Henau S, Feinstein J, Miller SI, Han B, Frokjaer-Jensen C, 
            Griffin EE
  TITLE     SapTrap Assembly of Caenorhabditis elegans MosSCI Transgene Vectors.
  JOURNAL   G3 (Bethesda). 2019 Dec 17. pii: g3.119.400822. doi: 
            10.1534/g3.119.400822.
  PUBMED    31848219
REFERENCE   2  (bases 1 to 4318)
  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: "G3 
            (Bethesda). 2019 Dec 17. pii: g3.119.400822. doi: 
            10.1534/g3.119.400822."
COMMENT     Sequence Label: pSDH66
FEATURES             Location/Qualifiers
     source          1..4318
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        239..257
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     239..256
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1206..1225)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1207..1225)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1232..1249)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1232..1248)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1241..1263)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             1386..1688
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     1567..1586
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             2037..2831
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             2037..2831
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2091..2110)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2701..2720
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     regulatory      3031..3040
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3037..3411
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     rep_origin      3549..4137
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4038..4057
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4291..4308
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctat
      241 ttaggtgaca ctatagaata ctcaagctat gcatcaagct tggtaccgag ctcggatcca
      301 ctagtaacgg ccgccagtgt gctggaattc aggcctgaat tcgcccttgc agctcttcgt
      361 acaaacagtt atgtttggta tattgggaat gtattctgtc atttaaggca aaaaccccat
      421 agacactact ccactttcaa attatcaact tctactcacc ttccactgag cctcaaaccc
      481 aaaccttctt ccgatctttc ttgcatcgtg ctcatcaata cttgtgattt tagcaagaaa
      541 aacgccgtcc tcgaaatctt gacaaacaat aggggaaacc ccaaaaagca aaagcaggaa
      601 aattaatttc ataaactttc acttgattga aaaattccaa aaatactcca aaactttttc
      661 acgatgagca tgatttgacg tcatgagagg cccagacgtg cgaagaaata aaaattgcgg
      721 tcataaactg aaacgtaaca tatgataagg tattttgtgt gcggtttttt ctatgatgtt
      781 tcaaaaaaat ttatcagaag taaaaaacat aagaagtgta cacagaacat tgtgtagatt
      841 ttgggataga attattatta atggggggag ggagcacaat ttttttcgac tttttattac
      901 gaattaattc tatttttgaa tctacacaat ttcattgtta gaggtgactt aaaagaagct
      961 aaaaaacaaa gaaattaaga gaagtttttt gataataaca aaaatagggg gtgggagcac
     1021 agggagaaag agcatgtagg gatgttgaag agtaattgga cttagaagtc agaggcacgg
     1081 gcgcgagatg gcgatctgat gacagcggcc gatgcggagc tcttacgaag agctgcaagg
     1141 gcgaattctg cagatatcca tcacactggc ggccgctcga gcatgcatct agagggccca
     1201 attcgcccta tagtgagtcg tattacaatt cactggccgt cgttttacaa cgtcgtgact
     1261 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     1321 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctatacg
     1381 tacggcagtt taaggtttac acctataaaa gagagagccg ttatcgtctg tttgtggatg
     1441 tacagagtga tattattgac acgccggggc gacggatggt gatccccctg gccagtgcac
     1501 gtctgctgtc agataaagtc tcccgtgaac tttacccggt ggtgcatatc ggggatgaaa
     1561 gctggcgcat gatgaccacc gatatggcca gtgtgccggt ctccgttatc ggggaagaag
     1621 tggctgatct cagccaccgc gaaaatgaca tcaaaaacgc cattaacctg atgttctggg
     1681 gaatataaat gtcaggcatg agattatcaa aaaggatctt cacctagatc cttttcacgt
     1741 agaaagccag tccgcagaaa cggtgctgac cccggatgaa tgtcagctac tgggctatct
     1801 ggacaaggga aaacgcaagc gcaaagagaa agcaggtagc ttgcagtggg cttacatggc
     1861 gatagctaga ctgggcggtt ttatggacag caagcgaacc ggaattgcca gctggggcgc
     1921 cctctggtaa ggttgggaag ccctgcaaag taaactggat ggctttcttg ccgccaagga
     1981 tctgatggcg caggggatca agctctgatc aagagacagg atgaggatcg tttcgcatga
     2041 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     2101 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     2161 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag
     2221 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     2281 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     2341 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     2401 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     2461 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     2521 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg
     2581 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     2641 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     2701 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     2761 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     2821 agttcttctg aattattaac gcttacaatt tcctgatgcg gtattttctc cttacgcatc
     2881 tgtgcggtat ttcacaccgc atcaggtggc acttttcggg gaaatgtgcg cggaacccct
     2941 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     3001 taaatgcttc aataatagca cgtgaggagg gccaccatgg ccaagttgac cagtgccgtt
     3061 ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt tctggaccga ccggctcggg
     3121 ttctcccggg acttcgtgga ggacgacttc gccggtgtgg tccgggacga cgtgaccctg
     3181 ttcatcagcg cggtccagga ccaggtggtg ccggacaaca ccctggcctg ggtgtgggtg
     3241 cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg tgtccacgaa cttccgggac
     3301 gcctccgggc cggccatgac cgagatcggc gagcagccgt gggggcggga gttcgccctg
     3361 cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg agcaggactg acacgtgcta
     3421 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     3481 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     3541 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     3601 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     3661 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     3721 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     3781 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     3841 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     3901 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     3961 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     4021 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     4081 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     4141 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     4201 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     4261 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaag
//
