LOCUS       sequence-335016-        2241 bp    DNA     circular SYN 15-APR-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) uses 
            blue-white screening.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2241)
  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 2241)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-Omega2
FEATURES             Location/Qualifiers
     source          1..2241
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(15..42)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     183..200
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     184..200
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     207..226
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        207..225
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        260..307
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     primer_bind     complement(688..707)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        complement(754..772)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(755..772)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(790..806)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(790..806)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      877..949
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      1005..1361
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             complement(1475..2134)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2049..2068
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2135..2237)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
ORIGIN
        1 attgtagaaa cgcaaaaagg ccatccgtca ggatggcctt ctgcttaatt tgatgcctgg
       61 cagtttatgg cgggcgtcct gcccgccacc ctttcagcca aaaaacttaa gaccgccggt
      121 cttgtccact accttgcagt aatgcggtgg acaggatcgg cggttttctt ttctcttctc
      181 aatgtaaaac gacggccagt gaattgtaat acgactcact atagggttcc tgcaggtctc
      241 agtcaggaga gagacgcctg ttgacaatta atcatcggca tagtatatcg gcatagtata
      301 atacgacaag gtgaggaact aaacccagga ggcagagatc atgaccatga ttacgccaag
      361 ctatctgggc gatactatcg aatactctag cttgcatgcg tgcaggtcga ctctagaaga
      421 tccccgggta ccgagctcta attcgccgta cagcgagtct tactacaatt ctctggcggt
      481 ggtcttgcaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
      541 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
      601 acagttgcgc agcctgaatg gcgaatgacg cctgatgcgg tattttctcc ttacgttact
      661 gtgcggtatt tcacacgtgc acctcagtac aatctgctct gatgccgcat agcgtctctc
      721 gcttgagacc tgcagtgact tgtattccgc ggttctatag tgtcacctaa atagcttggc
      781 gtaatttagg tcatagctgt ttcctgtcgg accaaaacga aaaaaggccc ccctttcggg
      841 aggcctcttt tctggaattt ggtaccgagc aggcatcaaa taaaacgaaa ggctcagtcg
      901 aaagactggg cctttcgttt tatctgttgt ttgtcggttg aacgctctct gacgtaccgc
      961 ggccgcgtag aggatctgtt gatcagcagt tcaacctgtt gatagtacgt actaagctct
     1021 catgtttcac gtactaagct ctcatgttta acgtactaag ctctcatgtt taacgaacta
     1081 aaccctcatg gctaacgtac taagctctca tggctaacgt actaagctct catgtttcac
     1141 gtactaagct ctcatgtttg aacaataaaa ttaatataaa tcagcaactt aaatagcctc
     1201 taaggtttta agttttataa gaaaaaaaag aatatataag gcttttaaag cttttaaggt
     1261 ttaacggttg tggacaacaa gccagggatg taacgcactg agaagccctt agagcctctc
     1321 aaagcaattt tgagtgacac aggaacactt aacggctgac atgtgatcat gtttgacagc
     1381 ttatcatcga atttctgcca ttcatccgct tattatcact tattcaggcg tagcaaccag
     1441 gcgtttaagg gcaccaataa ctgccttaaa aaaattacgc cccgccctgc cactcgtcgc
     1501 aatactgttg taattcatta agcattctgc cgacatggaa gccatcacaa acggcatgat
     1561 gaacctgaat cgccagcggc atcagcacct tgtcgccttg cgtatagtat ttgcccatcg
     1621 tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa ctggtgaaac
     1681 tcacccaggg attggctgac acgaaaaaca tattctcaat aaacccttta gggaaatagg
     1741 ccaggttttc accgtaacac gccacatctt gcgaatatat gtgtagaaac tgccggaaat
     1801 cgtcgtggta ttcactccag aggcttgaaa acgtttcagt ttgctcatgg aaaacggtgt
     1861 aacaagggtg aacactatcc catatcacca gctcaccgtc tttcattgcc atacgaaatt
     1921 ccggatgagc attcatcagg cgggcaagaa tgtgaataaa ggccggataa aacttgtgct
     1981 tatttttctt tacggtcttt aaaaaggccg taatatccag ttgaacggtc tggttatagg
     2041 tacattgagc aactgactga aatgcctcaa aatgttcttt acgatgccat tgggatatat
     2101 caacggtggt atatccagtg atttttttct ccattttagc ttccttagct cctgaaaatc
     2161 tcgataactc aaaaaatacg cccggtagtg atcttatttc attatggtga aagttggaac
     2221 ctcttacgtg ccgatcataa c
//
