LOCUS       sequence-309586-        4170 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MosSCI SapTrap assembly: Cassette 2 for 4- or 5-cassette system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4170)
  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 4170)
  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: pXF262
FEATURES             Location/Qualifiers
     source          1..4170
                     /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(1058..1077)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1059..1077)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1084..1101)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1084..1100)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1093..1115)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             1238..1540
                     /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     1419..1438
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             1889..2683
                     /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             1889..2683
                     /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(1943..1962)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2553..2572
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     regulatory      2883..2892
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2889..3263
                     /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      3401..3989
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3890..3909
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4143..4160
                     /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 gcccttgcag ctcttcgacc cttccacttc
      361 ttgttctcgt aatcgtagtg tgtcggtgca ccgaggaatt gtcccttctc gagcacaagc
      421 gcacgctcaa cctgagcctc cttgtactcg ttctggaagg ttggcggcag ttctgggtac
      481 acgcagacgt tgagaagaac agcataatag gtgcaaagtc cgagcacgga gaggataacg
      541 gcggagatca ccttccagta tccggttgga gcctcgaatt cggacagcgt ttgacggaag
      601 ctgtagcggt acactgaaaa cttagcaaca ttttcaagtt tgtcgaggaa aattgaatac
      661 taacagagct tcttctcctc agcagacaag ttcttccagt cggcttgctc cttggcgcga
      721 attggggcaa tcacactgtc ctccttgcgg aatcgaatag ctggatacca gtagtcgaga
      781 cgatcttggt agatctgaaa aataaattaa ttaaatttaa aatttataaa ataaacaagg
      841 cgtacgttat ctccatttgc tccgtatccg acgagctcgc gtccagcggc cttatcgggt
      901 ccccagtaga attcctgtcc tccggacaga cgggcggtgg tgtgcactgc gcggagagcc
      961 aaacgtggca gcatcatcga agagctgcaa gggcgaattc tgcagatatc catcacactg
     1021 gcggccgctc gagcatgcat ctagagggcc caattcgccc tatagtgagt cgtattacaa
     1081 ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
     1141 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
     1201 tcgcccttcc caacagttgc gcagcctata cgtacggcag tttaaggttt acacctataa
     1261 aagagagagc cgttatcgtc tgtttgtgga tgtacagagt gatattattg acacgccggg
     1321 gcgacggatg gtgatccccc tggccagtgc acgtctgctg tcagataaag tctcccgtga
     1381 actttacccg gtggtgcata tcggggatga aagctggcgc atgatgacca ccgatatggc
     1441 cagtgtgccg gtctccgtta tcggggaaga agtggctgat ctcagccacc gcgaaaatga
     1501 catcaaaaac gccattaacc tgatgttctg gggaatataa atgtcaggca tgagattatc
     1561 aaaaaggatc ttcacctaga tccttttcac gtagaaagcc agtccgcaga aacggtgctg
     1621 accccggatg aatgtcagct actgggctat ctggacaagg gaaaacgcaa gcgcaaagag
     1681 aaagcaggta gcttgcagtg ggcttacatg gcgatagcta gactgggcgg ttttatggac
     1741 agcaagcgaa ccggaattgc cagctggggc gccctctggt aaggttggga agccctgcaa
     1801 agtaaactgg atggctttct tgccgccaag gatctgatgg cgcaggggat caagctctga
     1861 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     1921 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     1981 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     2041 cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc
     2101 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     2161 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     2221 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     2281 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     2341 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     2401 cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     2461 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     2521 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     2581 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     2641 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgaattatta acgcttacaa
     2701 tttcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatcaggtg
     2761 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     2821 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatag cacgtgagga
     2881 gggccaccat ggccaagttg accagtgccg ttccggtgct caccgcgcgc gacgtcgccg
     2941 gagcggtcga gttctggacc gaccggctcg ggttctcccg ggacttcgtg gaggacgact
     3001 tcgccggtgt ggtccgggac gacgtgaccc tgttcatcag cgcggtccag gaccaggtgg
     3061 tgccggacaa caccctggcc tgggtgtggg tgcgcggcct ggacgagctg tacgccgagt
     3121 ggtcggaggt cgtgtccacg aacttccggg acgcctccgg gccggccatg accgagatcg
     3181 gcgagcagcc gtgggggcgg gagttcgccc tgcgcgaccc ggccggcaac tgcgtgcact
     3241 tcgtggccga ggagcaggac tgacacgtgc taaaacttca tttttaattt aaaaggatct
     3301 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     3361 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     3421 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     3481 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     3541 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     3601 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     3661 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     3721 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     3781 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     3841 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     3901 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     3961 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     4021 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     4081 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     4141 gcagcgagtc agtgagcgag gaagcggaag
//
