LOCUS       sequence-309583-        4380 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 4380)
  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 4380)
  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: pSM12
FEATURES             Location/Qualifiers
     source          1..4380
                     /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(1268..1287)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1269..1287)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1294..1311)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1294..1310)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1303..1325)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             1448..1750
                     /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     1629..1648
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             2099..2893
                     /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             2099..2893
                     /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(2153..2172)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2763..2782
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     regulatory      3093..3102
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3099..3473
                     /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      3611..4199
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4100..4119
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4353..4370
                     /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 gccgcttccc
      361 catacttgac ttggtagagg actgtatctt gcaactgcgt gttcgtatag cttaactttg
      421 ttgtaatccg aatcgtttcc tagtatttca attctgtgat caacaaagtg tgcatctgga
      481 agttggacga ccttctttgc tttgtaagtg gttttaaaat cgcaaaggta gtgacctcca
      541 ccttctagta gtagtgccat gttaatgttc tgaaaattta aatataatcc gttagttagt
      601 aagtccaagt ttaaacttac tccaacaagt agtccatctc taacatgtag tttttcagtc
      661 gatggttccc acttaagagt cttcttttgc ataactggtc cgttaggagg aaagttagta
      721 cctttaaatc tgacattttg gaagaagcaa tcaccttcga gactaatatc acttctctga
      781 aaatttaaat atgtatggtt agttagtacc gaactgttta aacttacaat agtgcaaata
      841 cctttatctt cgaaagtcat agttctttcc cacgagtatc cttctggaaa cgattgcttg
      901 aagtaatcag gaatatcttc tgggtattta gtaaaaactc tgtttccgta atggacggca
      961 gtagtgagaa tatcctgaaa atttaaataa tcagggttag ttagtatata tatgtttaaa
     1021 cttacgtaac taaatggtag tggtgctcct tctttgacag tcaagttagc agtttgagtt
     1081 ccttcgtatg gctttccttt accttcacct tcaataacaa atgcatgacc gttgacgttt
     1141 ccttccatat gcactttgac tctcatatct tccttaataa ggttcatcga agagctgcaa
     1201 gggcgaattc tgcagatatc catcacactg gcggccgctc gagcatgcat ctagagggcc
     1261 caattcgccc tatagtgagt cgtattacaa ttcactggcc gtcgttttac aacgtcgtga
     1321 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     1381 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctata
     1441 cgtacggcag tttaaggttt acacctataa aagagagagc cgttatcgtc tgtttgtgga
     1501 tgtacagagt gatattattg acacgccggg gcgacggatg gtgatccccc tggccagtgc
     1561 acgtctgctg tcagataaag tctcccgtga actttacccg gtggtgcata tcggggatga
     1621 aagctggcgc atgatgacca ccgatatggc cagtgtgccg gtctccgtta tcggggaaga
     1681 agtggctgat ctcagccacc gcgaaaatga catcaaaaac gccattaacc tgatgttctg
     1741 gggaatataa atgtcaggca tgagattatc aaaaaggatc ttcacctaga tccttttcac
     1801 gtagaaagcc agtccgcaga aacggtgctg accccggatg aatgtcagct actgggctat
     1861 ctggacaagg gaaaacgcaa gcgcaaagag aaagcaggta gcttgcagtg ggcttacatg
     1921 gcgatagcta gactgggcgg ttttatggac agcaagcgaa ccggaattgc cagctggggc
     1981 gccctctggt aaggttggga agccctgcaa agtaaactgg atggctttct tgccgccaag
     2041 gatctgatgg cgcaggggat caagctctga tcaagagaca ggatgaggat cgtttcgcat
     2101 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     2161 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     2221 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     2281 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     2341 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     2401 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     2461 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     2521 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     2581 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg
     2641 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     2701 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     2761 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     2821 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     2881 cgagttcttc tgaattatta acgcttacaa tttcctgatg cggtattttc tccttacgca
     2941 tctgtgcggt atttcacacc gcatcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     3001 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     3061 gataaatgct tcaataatag cacgtgagga gggccaccat ggccaagttg accagtgccg
     3121 ttccggtgct caccgcgcgc gacgtcgccg gagcggtcga gttctggacc gaccggctcg
     3181 ggttctcccg ggacttcgtg gaggacgact tcgccggtgt ggtccgggac gacgtgaccc
     3241 tgttcatcag cgcggtccag gaccaggtgg tgccggacaa caccctggcc tgggtgtggg
     3301 tgcgcggcct ggacgagctg tacgccgagt ggtcggaggt cgtgtccacg aacttccggg
     3361 acgcctccgg gccggccatg accgagatcg gcgagcagcc gtgggggcgg gagttcgccc
     3421 tgcgcgaccc ggccggcaac tgcgtgcact tcgtggccga ggagcaggac tgacacgtgc
     3481 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     3541 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     3601 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     3661 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     3721 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     3781 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     3841 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     3901 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     3961 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     4021 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     4081 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     4141 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     4201 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     4261 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     4321 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
//
