LOCUS       sequence-288131-        6156 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MosSCI SapTrap assembly: Cassette 3A for 5-cassette system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6156)
  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 6156)
  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: pSDH51
FEATURES             Location/Qualifiers
     source          1..6156
                     /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(3044..3063)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3045..3063)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3070..3087)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3070..3086)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3079..3101)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             3224..3526
                     /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     3405..3424
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             3875..4669
                     /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             3875..4669
                     /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(3929..3948)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4539..4558
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     regulatory      4869..4878
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4875..5249
                     /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      5387..5975
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5876..5895
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6129..6146
                     /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 ctcttcgggt ggaagcggcg
      361 ccgaggtaac acttagtttt tgttgctctg tttaaaaatt aattttaaga tcggaaccgg
      421 attcccattc gacccacact acgtcgaggt cctcggagag cgtatgcact acgtcgacgt
      481 cggaccacgt gacggaaccc cagtcctctt cctccacgga aacccaacct cctcctacgt
      541 ctggcgtaac atcatcccac acgtcgcccc aacccaccgt tgcatcgccc caggtaagtt
      601 ttcttatggg aaagaaggaa aaaaccgaga ttttacttga aaaattgaat ttttcgcggg
      661 attttcacca aaaattgttg aatattcatt atttcacgct gtaaaacaaa aaaaaaaaaa
      721 atcaaaaact acgttgaaat cgcgttttta agcgaatttt cttcagaatt gccagatttt
      781 aaccccaaat tttgcagttt ttaaataaaa tttcaccttt tcggctcaaa ttgtagattt
      841 tcctgaaaat ttagtacaaa aaacaatttc ctcgtaaatt tttcaaatag attttcagac
      901 ctcatcggaa tgggaaagtc cgacaagcca gacctcggat acttcttcga cgaccacgtc
      961 cgtttcatgg acgccttcat cgaggccctc ggactcgagg aggtcgtcct cgtcatccac
     1021 gactggggat cggctctcgg attccactgg gccaagcgta acccagagcg tgtcaaggga
     1081 atcgccttca tggagttcat ccgtccaatc ccaacctggg acgagtggcc aggtaagtta
     1141 tgtctacctg cctgcctacc gcctaaattt tttgtgaagt ttcttcaaaa aatccagaaa
     1201 aaaaaacaat tttcatacga ttttttccct taaaattgtg aattttcatg ctttttagcc
     1261 ccaaaagtca ttatttgaga aaaaatttca tacaaaaaag ttttgagaaa tacacaattt
     1321 tttaaatgta attttcaaat tttcaatttt caactagaaa attcacaaaa cttgtaaatt
     1381 ttggaccaaa acatttatac aattactttt tttgaatcta ataactacaa taactacaaa
     1441 tttaacattt tcagagttcg cccgtgagac attccaagcc ttccgtacca ccgacgtcgg
     1501 acgtaagctc atcatcgacc aaaacgtctt catcgaggga accctcccaa tgggagtcgt
     1561 ccgtccactc accgaggtcg agatggacca ctaccgtgag ccattcctca acccagtcga
     1621 ccgtgagcca ctctggcgtt tcccaaacga gctcccaatc gccggagagc cagccaacat
     1681 cgtcgccctc gtcgaggagt acatggactg gctccaccaa tccccagtcc caaagctcct
     1741 cttctgggga accccaggta agttcttttt ttgaaaagtg cagtttgtag tctaattttc
     1801 atttattttc tttttaaaaa acgcatcaat ttttaaatat tttgggacaa aatccgaaaa
     1861 ctgtactaat ttgtagtttg taaaattaaa aaaaaacgca aaaaaatgtt ttcaaaaatg
     1921 cttagaataa aaataaaatt tctaaaaaat tgaacaaaat aaacatttaa aaattcaaaa
     1981 agtttgaaaa aatgcaaagt tttttaatgc aaaaaatatt taatgtttaa acaaatttaa
     2041 aaaatgttgt aagaatatgt ttagagacag tttttaaatg ttaaattgca aaaaaaaaaa
     2101 gcaaaaaaaa atttgaaaaa tgtaacagaa aattttaatt aaaaaagtcg tgtttttttc
     2161 aaacctgcaa ttctttgtaa ttttatttat aaagagaaac aaatcaaatt gtatattacg
     2221 aaaaacgata ctaaaattcg aaaatttgcg gttttttgcg taaaaaatac ggttccgtaa
     2281 ttttcaggag tcctcatccc accagccgag gccgcccgtc tcgccaagtc cctcccaaac
     2341 tgcaaggccg tcgacatcgg accaggactc aacctcctcc aagaggacaa cccaggtaag
     2401 ttcatagatt ttgaaaaaaa gtttaagaac tgaaaaatgg aataaaaata tttaagagca
     2461 tttttaaatg taaaattaac aaaaaaagcg cctaagaatg tttccaaaac agtaaaaaaa
     2521 aggtttaaaa aaatgcaaaa aaaaatttaa gatgctttta attacacaaa aaatgaaagt
     2581 aaaaaaaatt aaaatattga aaaatgtata tttgtttgaa aaggtacatt tttcaatgca
     2641 aaaaatgttt ctaaaaatta aaaaaatatg tttaaattgt gtaaagtgta aattaaaaaa
     2701 taaaattatg aaaaagtatt gtttcatact ttttctaaat tttggttgaa aaatttaaaa
     2761 ttccagcttt ttattcacat tatgtgttct aaaaatcttt ccccaatttt cgggaccgtt
     2821 ttccagtata tttccatttt tcaaaaagat ttttaactga aattcatgtt ttcaatgcta
     2881 aaaatcaata aaaaaagaat ttttcagacc tcatcggttc cgagatcgcc cgttggctct
     2941 ccaccctcga gatctccgga acgcgaagag ctgcaagggc gaattctgca gatatccatc
     3001 acactggcgg ccgctcgagc atgcatctag agggcccaat tcgccctata gtgagtcgta
     3061 ttacaattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
     3121 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
     3181 caccgatcgc ccttcccaac agttgcgcag cctatacgta cggcagttta aggtttacac
     3241 ctataaaaga gagagccgtt atcgtctgtt tgtggatgta cagagtgata ttattgacac
     3301 gccggggcga cggatggtga tccccctggc cagtgcacgt ctgctgtcag ataaagtctc
     3361 ccgtgaactt tacccggtgg tgcatatcgg ggatgaaagc tggcgcatga tgaccaccga
     3421 tatggccagt gtgccggtct ccgttatcgg ggaagaagtg gctgatctca gccaccgcga
     3481 aaatgacatc aaaaacgcca ttaacctgat gttctgggga atataaatgt caggcatgag
     3541 attatcaaaa aggatcttca cctagatcct tttcacgtag aaagccagtc cgcagaaacg
     3601 gtgctgaccc cggatgaatg tcagctactg ggctatctgg acaagggaaa acgcaagcgc
     3661 aaagagaaag caggtagctt gcagtgggct tacatggcga tagctagact gggcggtttt
     3721 atggacagca agcgaaccgg aattgccagc tggggcgccc tctggtaagg ttgggaagcc
     3781 ctgcaaagta aactggatgg ctttcttgcc gccaaggatc tgatggcgca ggggatcaag
     3841 ctctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     3901 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     3961 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     4021 caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg
     4081 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     4141 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     4201 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     4261 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     4321 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     4381 actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg
     4441 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     4501 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     4561 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     4621 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgaa ttattaacgc
     4681 ttacaatttc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     4741 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     4801 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatagcacg
     4861 tgaggagggc caccatggcc aagttgacca gtgccgttcc ggtgctcacc gcgcgcgacg
     4921 tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac ttcgtggagg
     4981 acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg gtccaggacc
     5041 aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg cggcctggac gagctgtacg
     5101 ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc ctccgggccg gccatgaccg
     5161 agatcggcga gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc ggcaactgcg
     5221 tgcacttcgt ggccgaggag caggactgac acgtgctaaa acttcatttt taatttaaaa
     5281 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     5341 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     5401 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     5461 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     5521 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     5581 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     5641 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     5701 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     5761 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     5821 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     5881 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     5941 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     6001 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     6061 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     6121 ccgagcgcag cgagtcagtg agcgaggaag cggaag
//
