LOCUS       sequence-273937-        4378 bp    DNA     circular SYN 14-APR-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 4378)
  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 4378)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: pSDH61
FEATURES             Location/Qualifiers
     source          1..4378
                     /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(1266..1285)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1267..1285)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1292..1309)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1292..1308)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1301..1323)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             1446..1748
                     /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     1627..1646
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             2097..2891
                     /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             2097..2891
                     /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(2151..2170)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2761..2780
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     regulatory      3091..3100
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3097..3471
                     /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      3609..4197
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4098..4117
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4351..4368
                     /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 agctcccgtc
      361 gcgacgggtg gatcacgggc gcgtggcact gtcgattgca aaatacggag cttgagctcc
      421 aatctactcg ttctataatt gatggagatt ggtgatgagt agtaatgata gttgtaatga
      481 gcatagacag tgatggtgta gtcgactcct ggcttcaatc cggaaattgt ggcggtcgac
      541 ttgcttcctg aaaatattgt actctccaaa ttacttttag attctcataa aaattaatta
      601 acttacctgg gaccgtgaat tcttggactg gagaatttcc tccggtctct ccgtaggtga
      661 tcctatagta ggagactgga agttcacggt aagcatccca ggagatgaga aggcttgttg
      721 gtgtggcagc aacgacctca aggttggttg gaacactgga aactcctgaa aaattagaaa
      781 aagtaagtca gttttaaatt tgtaggagaa attataactt acctccggat cctccgtcct
      841 tttcgacgcg gacacggatc tcctgcttag ctccgtttcc tacctctcgg acgatgatga
      901 gctcgactgt gttctggaag gtcttgagaa ggctgacagc ctgaccgtgc tcaatattaa
      961 tgaaagagta accgttggct tggatgatct gaaaagtctt catgaatgag aaattgcacg
     1021 atctctcgat gaaattaatt aacttacctt gtctccaggt tgaagaagtt tgctagctgg
     1081 tccttctggt tggacccgag taacaaaaat tccatcatcg tcaggacgga atggatttcc
     1141 acggcctccg acacctccgg agatcgagaa tccgagctct ggcatcgaag agctgcaagg
     1201 gcgaattctg cagatatcca tcacactggc ggccgctcga gcatgcatct agagggccca
     1261 attcgcccta tagtgagtcg tattacaatt cactggccgt cgttttacaa cgtcgtgact
     1321 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     1381 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctatacg
     1441 tacggcagtt taaggtttac acctataaaa gagagagccg ttatcgtctg tttgtggatg
     1501 tacagagtga tattattgac acgccggggc gacggatggt gatccccctg gccagtgcac
     1561 gtctgctgtc agataaagtc tcccgtgaac tttacccggt ggtgcatatc ggggatgaaa
     1621 gctggcgcat gatgaccacc gatatggcca gtgtgccggt ctccgttatc ggggaagaag
     1681 tggctgatct cagccaccgc gaaaatgaca tcaaaaacgc cattaacctg atgttctggg
     1741 gaatataaat gtcaggcatg agattatcaa aaaggatctt cacctagatc cttttcacgt
     1801 agaaagccag tccgcagaaa cggtgctgac cccggatgaa tgtcagctac tgggctatct
     1861 ggacaaggga aaacgcaagc gcaaagagaa agcaggtagc ttgcagtggg cttacatggc
     1921 gatagctaga ctgggcggtt ttatggacag caagcgaacc ggaattgcca gctggggcgc
     1981 cctctggtaa ggttgggaag ccctgcaaag taaactggat ggctttcttg ccgccaagga
     2041 tctgatggcg caggggatca agctctgatc aagagacagg atgaggatcg tttcgcatga
     2101 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     2161 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     2221 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag
     2281 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     2341 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     2401 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     2461 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     2521 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     2581 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg
     2641 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     2701 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     2761 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     2821 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     2881 agttcttctg aattattaac gcttacaatt tcctgatgcg gtattttctc cttacgcatc
     2941 tgtgcggtat ttcacaccgc atcaggtggc acttttcggg gaaatgtgcg cggaacccct
     3001 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     3061 taaatgcttc aataatagca cgtgaggagg gccaccatgg ccaagttgac cagtgccgtt
     3121 ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt tctggaccga ccggctcggg
     3181 ttctcccggg acttcgtgga ggacgacttc gccggtgtgg tccgggacga cgtgaccctg
     3241 ttcatcagcg cggtccagga ccaggtggtg ccggacaaca ccctggcctg ggtgtgggtg
     3301 cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg tgtccacgaa cttccgggac
     3361 gcctccgggc cggccatgac cgagatcggc gagcagccgt gggggcggga gttcgccctg
     3421 cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg agcaggactg acacgtgcta
     3481 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     3541 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     3601 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     3661 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     3721 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     3781 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     3841 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     3901 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     3961 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     4021 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     4081 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     4141 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     4201 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     4261 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     4321 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaag
//
