LOCUS       sequence-273920-        4527 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 4527)
  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 4527)
  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: pJF7
FEATURES             Location/Qualifiers
     source          1..4527
                     /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(1415..1434)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1416..1434)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1441..1458)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1441..1457)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1450..1472)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             1595..1897
                     /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     1776..1795
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             2246..3040
                     /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             2246..3040
                     /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(2300..2319)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2910..2929
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     regulatory      3240..3249
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3246..3620
                     /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      3758..4346
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4247..4266
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4500..4517
                     /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 ctccaagtcg
      361 tcggtgtcat cgataagctc ttggatttgt gctggtggag gtccgtctcc gaggtattga
      421 accattggtg gctcgttttc tgcctcatct gctttcgctc cgaacatttc cgcaagacgc
      481 taaaaattaa catagaagat attgaaaatt gaaaaaataa accaatttct gtgagtaaaa
      541 aattctagtt tgaatatatg cctacttaac tagaattttg aataagttct cctagatatt
      601 cgaatgttta atttgcattt gaaactgttg tactgaaata acgcattttt gatggaatgt
      661 ggctcaaaat tataagaaaa ttttgcatcc ttcaaaattg tatatttaat ttcgaatatt
      721 ttctcggaca ggttaatttg aatgccaggt tttcacatga aaatttttta aatagtattt
      781 aacagtcaca tttcaataac ttttaaagtg aatttttaca tttaaaaacc acttacttcg
      841 cgagtcgacg gcaattgttg aacaacagca cggaactgat cttccgacaa tgtttgttgg
      901 aagatagaca ggagttgttg agattcaccg cagagcatga ctgcgttggc aatgtgcacg
      961 gcaccttcat caacatttcc ggctgccatg agttcttctc cgagttgaac ctcttgcaag
     1021 aagaaccttt gcatttggga tggatcggtg acatctggag cagcatcatt gcctgctgca
     1081 gctggacgac gtggagccat tcctcctgct ccagcctggg cacgtctctc tgaaagaaaa
     1141 taagttgttt ttagataaaa atccgaagga aatatgttgt taaaacactg actttgccta
     1201 atcttgtcct tgtagtctgg agcgttgatt ctcttatgat cgaagtaaat gcagtagccg
     1261 aggaaagcgg ctccagcaat tccagcagcc aaaacgacgt ttgatttgtt gaaaccaaga
     1321 attgtgtccg acatcgaaga gctgcaaggg cgaattctgc agatatccat cacactggcg
     1381 gccgctcgag catgcatcta gagggcccaa ttcgccctat agtgagtcgt attacaattc
     1441 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
     1501 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
     1561 cccttcccaa cagttgcgca gcctatacgt acggcagttt aaggtttaca cctataaaag
     1621 agagagccgt tatcgtctgt ttgtggatgt acagagtgat attattgaca cgccggggcg
     1681 acggatggtg atccccctgg ccagtgcacg tctgctgtca gataaagtct cccgtgaact
     1741 ttacccggtg gtgcatatcg gggatgaaag ctggcgcatg atgaccaccg atatggccag
     1801 tgtgccggtc tccgttatcg gggaagaagt ggctgatctc agccaccgcg aaaatgacat
     1861 caaaaacgcc attaacctga tgttctgggg aatataaatg tcaggcatga gattatcaaa
     1921 aaggatcttc acctagatcc ttttcacgta gaaagccagt ccgcagaaac ggtgctgacc
     1981 ccggatgaat gtcagctact gggctatctg gacaagggaa aacgcaagcg caaagagaaa
     2041 gcaggtagct tgcagtgggc ttacatggcg atagctagac tgggcggttt tatggacagc
     2101 aagcgaaccg gaattgccag ctggggcgcc ctctggtaag gttgggaagc cctgcaaagt
     2161 aaactggatg gctttcttgc cgccaaggat ctgatggcgc aggggatcaa gctctgatca
     2221 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     2281 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     2341 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     2401 cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac
     2461 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     2521 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     2581 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     2641 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     2701 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     2761 caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     2821 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     2881 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     2941 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     3001 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga attattaacg cttacaattt
     3061 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tcaggtggca
     3121 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     3181 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatagcac gtgaggaggg
     3241 ccaccatggc caagttgacc agtgccgttc cggtgctcac cgcgcgcgac gtcgccggag
     3301 cggtcgagtt ctggaccgac cggctcgggt tctcccggga cttcgtggag gacgacttcg
     3361 ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac caggtggtgc
     3421 cggacaacac cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac gccgagtggt
     3481 cggaggtcgt gtccacgaac ttccgggacg cctccgggcc ggccatgacc gagatcggcg
     3541 agcagccgtg ggggcgggag ttcgccctgc gcgacccggc cggcaactgc gtgcacttcg
     3601 tggccgagga gcaggactga cacgtgctaa aacttcattt ttaatttaaa aggatctagg
     3661 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     3721 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     3781 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     3841 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     3901 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     3961 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     4021 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     4081 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     4141 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     4201 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     4261 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     4321 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     4381 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     4441 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     4501 gcgagtcagt gagcgaggaa gcggaag
//
