LOCUS       sequence-275295-        4353 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 4353)
  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 4353)
  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: pSM10
FEATURES             Location/Qualifiers
     source          1..4353
                     /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(1241..1260)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1242..1260)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1267..1284)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1267..1283)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1276..1298)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             1421..1723
                     /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     1602..1621
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             2072..2866
                     /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             2072..2866
                     /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(2126..2145)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2736..2755
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     regulatory      3066..3075
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3072..3446
                     /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      3584..4172
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4073..4092
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4326..4343
                     /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 acctccacct
      361 cccttgtaca gctcgtccat tcctccggtg gagtggcgtc cctcggaacg ttcgtattgc
      421 tccacgacgg tgtagtcctc gttgtgggag gtgatgtcga gcttgcggtc gacgttgtag
      481 gctcctggca tctggactgg cttcttggcc ttgtaggtgg tcttgaagtc cgccaggtaa
      541 cgtcctccgt ccttcagacg gagggccatc ttaatgtctc cctgaaaatt taaatatgta
      601 tggttagtta gtaccgaact gtttaaactt accttgagca ctccgtcctc tgggtacaaa
      661 cgctcggtgg aggcttccca tcccatggtc ttcttctgca tgactgggcc gtcaggtggg
      721 aagttggttc cacggagctt caccttgtag atgagggttc cgtcctccag ggaggtgtct
      781 tgggtgacgg tcacggctcc tccgtcctcg aagttcatca cgcgctccca cttgaatccc
      841 tcggggaagg attgcttgta gtagtcgggg atgtcggctg ggtgcttggt gaaggcacgg
      901 gagccgtaca tgaattgtgg ggagaggatg tcccaggaga atggcagtgg tccaccctga
      961 aaatttaaat aatcagggtt agttagtata tatatgttta aacttacctt ggtcaccttg
     1021 agcttggcgg tctgggttcc ctcgtatggg cgtccctctc cctcgccctc gatctcgaac
     1081 tcgtgtccgt tcatggatcc ctccatgtgc accttgaaac gcatgaactc cttgatgact
     1141 gcctcgccct tggagaccat cgaagagctg caagggcgaa ttctgcagat atccatcaca
     1201 ctggcggccg ctcgagcatg catctagagg gcccaattcg ccctatagtg agtcgtatta
     1261 caattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
     1321 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
     1381 cgatcgccct tcccaacagt tgcgcagcct atacgtacgg cagtttaagg tttacaccta
     1441 taaaagagag agccgttatc gtctgtttgt ggatgtacag agtgatatta ttgacacgcc
     1501 ggggcgacgg atggtgatcc ccctggccag tgcacgtctg ctgtcagata aagtctcccg
     1561 tgaactttac ccggtggtgc atatcgggga tgaaagctgg cgcatgatga ccaccgatat
     1621 ggccagtgtg ccggtctccg ttatcgggga agaagtggct gatctcagcc accgcgaaaa
     1681 tgacatcaaa aacgccatta acctgatgtt ctggggaata taaatgtcag gcatgagatt
     1741 atcaaaaagg atcttcacct agatcctttt cacgtagaaa gccagtccgc agaaacggtg
     1801 ctgaccccgg atgaatgtca gctactgggc tatctggaca agggaaaacg caagcgcaaa
     1861 gagaaagcag gtagcttgca gtgggcttac atggcgatag ctagactggg cggttttatg
     1921 gacagcaagc gaaccggaat tgccagctgg ggcgccctct ggtaaggttg ggaagccctg
     1981 caaagtaaac tggatggctt tcttgccgcc aaggatctga tggcgcaggg gatcaagctc
     2041 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     2101 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     2161 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     2221 gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct
     2281 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     2341 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     2401 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     2461 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     2521 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     2581 gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     2641 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     2701 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     2761 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     2821 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgaatta ttaacgctta
     2881 caatttcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatcag
     2941 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     3001 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tagcacgtga
     3061 ggagggccac catggccaag ttgaccagtg ccgttccggt gctcaccgcg cgcgacgtcg
     3121 ccggagcggt cgagttctgg accgaccggc tcgggttctc ccgggacttc gtggaggacg
     3181 acttcgccgg tgtggtccgg gacgacgtga ccctgttcat cagcgcggtc caggaccagg
     3241 tggtgccgga caacaccctg gcctgggtgt gggtgcgcgg cctggacgag ctgtacgccg
     3301 agtggtcgga ggtcgtgtcc acgaacttcc gggacgcctc cgggccggcc atgaccgaga
     3361 tcggcgagca gccgtggggg cgggagttcg ccctgcgcga cccggccggc aactgcgtgc
     3421 acttcgtggc cgaggagcag gactgacacg tgctaaaact tcatttttaa tttaaaagga
     3481 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     3541 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     3601 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     3661 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     3721 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     3781 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     3841 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     3901 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     3961 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     4021 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     4081 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     4141 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     4201 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     4261 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     4321 agcgcagcga gtcagtgagc gaggaagcgg aag
//
