LOCUS       sequence-335014-        2243 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Omega level R6K_ origin GB Cloning vector, Conditionally replicative
            ori, requires pir gene expression requires pir gene expression 
            requires EC100D or EC100D116 bacterial strain (DH10B derived).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2243)
  AUTHORS   Matinyan N, Karkhanis MS, Gonzalez Y, Jain A, Saltzman A, 
            Malovannaya A, Sarrion-Perdigones A, Dierick HA, Venken KJT
  TITLE     Multiplexed drug-based selection and counterselection genetic 
            manipulations in Drosophila.
  JOURNAL   Cell Rep. 2021 Sep 14;36(11):109700. doi: 
            10.1016/j.celrep.2021.109700.
  PUBMED    34525356
REFERENCE   2  (bases 1 to 2243)
  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: "Cell Rep.";
            date: "2021-09-14"; volume: "36(11):109700. doi"; pages: " 
            10.1016/j.celrep.2021.109700"
COMMENT     Sequence Label: pR6Kg-Omega1
FEATURES             Location/Qualifiers
     source          1..2243
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(113..772)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     687..706
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(773..875)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(894..921)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     1062..1079
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1063..1079
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1086..1105
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1086..1104
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        1136..1183
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     primer_bind     complement(1564..1583)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        complement(1635..1653)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1636..1653)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1671..1687)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1671..1687)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      1758..1830
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      1886..2242
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
ORIGIN
        1 gtgatcatgt ttgacagctt atcatcgaat ttctgccatt catccgctta ttatcactta
       61 ttcaggcgta gcaaccaggc gtttaagggc accaataact gccttaaaaa aattacgccc
      121 cgccctgcca ctcgtcgcaa tactgttgta attcattaag cattctgccg acatggaagc
      181 catcacaaac ggcatgatga acctgaatcg ccagcggcat cagcaccttg tcgccttgcg
      241 tatagtattt gcccatcgtg aaaacggggg cgaagaagtt gtccatattg gccacgttta
      301 aatcaaaact ggtgaaactc acccagggat tggctgacac gaaaaacata ttctcaataa
      361 accctttagg gaaataggcc aggttttcac cgtaacacgc cacatcttgc gaatatatgt
      421 gtagaaactg ccggaaatcg tcgtggtatt cactccagag gcttgaaaac gtttcagttt
      481 gctcatggaa aacggtgtaa caagggtgaa cactatccca tatcaccagc tcaccgtctt
      541 tcattgccat acgaaattcc ggatgagcat tcatcaggcg ggcaagaatg tgaataaagg
      601 ccggataaaa cttgtgctta tttttcttta cggtctttaa aaaggccgta atatccagtt
      661 gaacggtctg gttataggta cattgagcaa ctgactgaaa tgcctcaaaa tgttctttac
      721 gatgccattg ggatatatca acggtggtat atccagtgat ttttttctcc attttagctt
      781 ccttagctcc tgaaaatctc gataactcaa aaaatacgcc cggtagtgat cttatttcat
      841 tatggtgaaa gttggaacct cttacgtgcc gatcataaca ttgtagaaac gcaaaaaggc
      901 catccgtcag gatggccttc tgcttaattt gatgcctggc agtttatggc gggcgtcctg
      961 cccgccaccc tttcagccaa aaaacttaag accgccggtc ttgtccacta ccttgcagta
     1021 atgcggtgga caggatcggc ggttttcttt tctcttctca atgtaaaacg acggccagtg
     1081 aattgtaata cgactcacta tagggttcgg atccggtctc aggagagaga cgcctgttga
     1141 caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac
     1201 ccaggaggca gagatcatga ccatgattac gccaagctat ctgggcgata ctatcgaata
     1261 ctctagcttg catgcgtgca ggtcgactct agaagatccc cgggtaccga gctctaattc
     1321 gccgtacagc gagtcttact acaattctct ggcggtggtc ttgcaacgtc gtgactggga
     1381 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     1441 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     1501 atgacgcctg atgcggtatt ttctccttac gttactgtgc ggtatttcac acgtgcacct
     1561 cagtacaatc tgctctgatg ccgcatagcg tctctcgctg tcatgagacc ggatcctgac
     1621 ttgtattccg cggttctata gtgtcaccta aatagcttgg cgtaatttag gtcatagctg
     1681 tttcctgtcg gaccaaaacg aaaaaaggcc cccctttcgg gaggcctctt ttctggaatt
     1741 tggtaccgag caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt
     1801 ttatctgttg tttgtcggtt gaacgctctc tgacgtaccg cggccgcgta gaggatctgt
     1861 tgatcagcag ttcaacctgt tgatagtacg tactaagctc tcatgtttca cgtactaagc
     1921 tctcatgttt aacgtactaa gctctcatgt ttaacgaact aaaccctcat ggctaacgta
     1981 ctaagctctc atggctaacg tactaagctc tcatgtttca cgtactaagc tctcatgttt
     2041 gaacaataaa attaatataa atcagcaact taaatagcct ctaaggtttt aagttttata
     2101 agaaaaaaaa gaatatataa ggcttttaaa gcttttaagg tttaacggtt gtggacaaca
     2161 agccagggat gtaacgcact gagaagccct tagagcctct caaagcaatt ttgagtgaca
     2221 caggaacact taacggctga cat
//
