LOCUS       sequence-342954-        3462 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Destination vector for Golden Gate Cloning assemblies, blue-white 
            screening.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3462)
  AUTHORS   Green EI, Jaouen E, Klug D, Proveti Olmo R, Gautier A, Blandin S, 
            Marois E
  TITLE     A population modification gene drive targeting both Saglin and 
            Lipophorin impairs Plasmodium transmission in Anopheles mosquitoes.
  JOURNAL   Elife. 2023 Dec 5;12:e93142. doi: 10.7554/eLife.93142.
  PUBMED    38051195
REFERENCE   2  (bases 1 to 3462)
  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: "Elife."; 
            date: "2023-12-5"; volume: "12:e93142. doi"; pages: " 
            10.7554/eLife.93142"
COMMENT     Sequence Label: pENTR R4 ATCC LacZ GCTT
FEATURES             Location/Qualifiers
     source          1..3462
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(603..727)
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    796..817
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(832..837,838..855,856..862)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    870..886
                     /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     875..897
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     894..910
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             906..1481
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWDAPCSGALSAAGVVVTRSVTATLASALAPAPFAFFPSFLATFAG
                     FPRQALNRGLPLGFRFSALRHLDPKKLD"
     primer_bind     929..949
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        931..949
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    962..1069
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     986..1002
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     986..1002
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(1036..1052)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(1078..1096)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1106..1122)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      1264..1644
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1351..1370)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1561..1582
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        complement(1704..1722)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1727..1743)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1845..1864)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1856..2665
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1923..1942)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2812..3400
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3301..3320
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccct acaggtcact aataccatct aagtagttga
      601 ttcatagtga ctggatatgt tgtgttttac agtattatgt agtctgtttt ttatgcaaaa
      661 tctaatttaa tatattgata tttatatcat tttacgtttc tcgttcaact tttctataca
      721 aagttggtac cgggcccccc gctagcgtcg acggtatcga taagcttgat cggatccaga
      781 gacccgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
      841 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
      901 cagctatgac catgattacg ccaagcgcgc aattaaccct cactaaaggg aacaaaagct
      961 ggagctccac cgcggtggcg gccgctctag aactagtgga tcccccgggc tgcaggaatt
     1021 cgatatcaag cttatcgata ccgtcgacct cgaggggggg cccggtaccc aattcgccct
     1081 atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa
     1141 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
     1201 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     1261 gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     1321 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     1381 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     1441 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     1501 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     1561 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     1621 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     1681 ttaacgcggg tctccgctta tcccctatag tgagtcgtat tacatggtca tagctgtttc
     1741 ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat
     1801 atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag
     1861 ccatattcaa cgggaaacgt cgaggccgcg attaaattcc aacatggatg ctgatttata
     1921 tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta
     1981 tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga
     2041 tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat
     2101 caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga tccccggaaa
     2161 aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct
     2221 ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga
     2281 tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag
     2341 tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa
     2401 acttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct
     2461 tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga
     2521 ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca
     2581 gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca
     2641 tttgatgctc gatgagtttt tctaatcaga attggttaat tggttgtaac actggcagag
     2701 cattacgctg acttgacggg acggcgcaag ctcatgacca aaatccctta acgtgagtta
     2761 cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     2821 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     2881 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     2941 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     3001 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     3061 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     3121 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     3181 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     3241 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     3301 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     3361 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     3421 tttacggttc ctggcctttt gctggccttt tgctcacatg tt
//
