LOCUS       sequence-345193-        3983 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Terminal pBlueScript derived high-copy GB Cloning vector with both 
            Alpha and Omega Entry points 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 3983)
  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 3983)
  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: pColE1-AlphaOmega
FEATURES             Location/Qualifiers
     source          1..3983
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             244..1035
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      1090..1545
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1177..1196)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1387..1408
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1569..1616
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        1670..1688
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1670..1687
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     misc_feature    1695..1751
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1753..1772)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1754..1772)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1779..1796)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1779..1795)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1788..1810)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2004..2023)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2063..2110
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     rep_origin      complement(3171..3759)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3251..3270)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgtaagctgt aatgcaagta gcgtatgcgc tcacgcaact ggtccagaac cttgaccgaa
       61 cgcagcggtg gtaacggcgc agtggcggtt ttcatggctt gttatgactg tttttttggg
      121 gtacagtcta tgcctcgggc atccaagcag caagcgcgtt acgccgtggg tcgatgtttg
      181 atgttatgga gcagcaacga tgttacgcag cagggcagtc gccctaaaac aaagttaaac
      241 attatgaggg aagcggtgat cgccgaagta tcgactcaac tatcagaggt agttggcgtc
      301 atcgagcgcc atctcgaacc gacgttgctg gccgtacatt tgtacggctc cgcagtggat
      361 ggcggcctga agccacacag tgatattgat ttgctggtta cggtgaccgt aaggcttgat
      421 gaaacaacgc ggcgagcttt gatcaacgac cttttggaaa cttcggcttc ccctggagag
      481 agcgagattc tccgcgctgt agaagtcacc attgttgtgc acgacgacat cattccgtgg
      541 cgttatccag ctaagcgcga actgcaattt ggagaatggc agcgcaatga cattcttgca
      601 ggtatcttcg agccagccac gatcgacatt gatctggcta tcttgctgac aaaagcaaga
      661 gaacatagcg ttgccttggt aggtccagcg gcggaggaac tctttgatcc ggttcctgaa
      721 caggatctat ttgaggcgct aaatgaaacc ttaacgctat ggaactcgcc gcccgactgg
      781 gctggcgatg agcgaaatgt agtgcttacg ttgtcccgca tttggtacag cgcagtaacc
      841 ggcaaaatcg cgccgaagga tgtagctgcc gactgggcaa tggagcgcct gccggcccag
      901 tatcagcccg tcatacttga agctagacag gcttatcttg gacaagaaga agatcgtttg
      961 gcctcgcgcg cagatcagtt ggaagaattt gtccactacg tgaaaggcga gatcaccaag
     1021 gtagtcggca aataatcact agaccaatgt tacacatata tactttagat tgatttaaaa
     1081 cttcatgtta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     1141 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     1201 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     1261 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     1321 gtagtgggcc agcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     1381 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     1441 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     1501 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aattttaacg gatcagcgag
     1561 agacccctgt tgacaattaa tcatcggcat agtatatcgg catagtataa tacgacaagg
     1621 tgaggaacta aacccaggag gcagatcatg accatgatta cgccaagcta tttaggtgac
     1681 actatagaat actcaagctt gcatgcctgc aggtcgactc tagaggatcc ccgggtaccg
     1741 agctcgaatt cgccctatag tgagtcgtat tacaattcac tggccgtcgt tttacaacgt
     1801 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     1861 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     1921 ctgaatggcg aatgacgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     1981 caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcataggg tctctctccg
     2041 gatcccaggg agagagacgc ctgttgacaa ttaatcatcg gcatagtata tcggcatagt
     2101 ataatacgac aaggtgagga actaaaccca ggaggcagat caatggcgag cttggttaag
     2161 aaagatatgt gtgttaagat gacgatggaa ggtactgtga acggttatca ctttaagtgc
     2221 gttggcgagg gtgaaggcaa gccgttcgag ggcacgcaga acatgcgcat tcgtgtcacc
     2281 gagggcggtc cgctgccttt tgcattcgac atcctggccc cgtgctgtat gtacggctct
     2341 aagaccttca ttaaacacgt gagcggtatc ccggattact ttaaagagtc ctttccagag
     2401 ggcttcactt gggaacgtac ccagattttt gaggacggtg gtgttctgac cgcgcaccaa
     2461 gacaccagcc tggaaggtaa ttgcctgatc tataaagtga aggttctggg taccaatttc
     2521 ccggcgaatg gtccggtgat gcaaaagaaa accgcgggtt gggagccgtg cgtcgagatg
     2581 ctgtatccgc gtgacggcgt cttgtgtggt cagagcttga tggcgctgaa gtgcaccgat
     2641 ggcaatcatc tgaccagcca cctgcgcacg acgtatcgta gccgtaaacc gagcaacgcc
     2701 gttaacatgc cggagttcca ttttggtgac catcgcatcg aaatcctgaa agctgagcag
     2761 ggcaaattct acgaacaata cgaatcggct gtcgcacgtt acagcgatgt gccggaaaaa
     2821 gcgacgtaat gagcttgaat tctgtacact cgagagtctc accccaaggg cgacacccca
     2881 taattagccc gggcgaaagg cccagtcttt cgactgagcc tttcgtttta tttgatgcct
     2941 ggcagttccc tactctcgca tggggagtcc ccacactacc atcggcgcta cggcgtttca
     3001 cttctgagtt cggcatgggg tcaggtggga ccaccgcgct actgccgcca ggcaaacgtc
     3061 tcacgcttga cacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     3121 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     3181 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     3241 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     3301 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     3361 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     3421 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     3481 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     3541 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     3601 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     3661 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     3721 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     3781 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     3841 tcaaaaagga tcttcaccta gatccttttg tgaccagcca ggacagaaat gcctcgactt
     3901 cgctgctacc caaggttgcc gggtgacgca caccgtggaa acggatgaag gcacgaaccc
     3961 agtggacata agcctgttcg gtt
//
