LOCUS       sequence-323403-        7529 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Hygromycin resistance vector with attB and eye marker. Contains an 
            Alpha GB cloning entry point for addition of custom assembly uses 
            blue-white screening.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7529)
  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 7529)
  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: pHygroR-GMRWhite
FEATURES             Location/Qualifiers
     source          1..7529
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(28..456)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(138..159)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     350..369
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(511..1302)
                     /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      1770..2358
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2259..2278
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2673..2807
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2710..2729)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2764..2783
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(5332..6357)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     promoter        complement(6447..6898)
                     /label=hsp70 promoter
                     /note="Drosophila melanogaster hsp70Bb promoter"
     primer_bind     complement(6478..6499)
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     protein_bind    6926..6995
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     primer_bind     7052..7071
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7265..7287
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7279..7296
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7280..7296
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7303..7322
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7303..7321
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    7324..7380
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     promoter        complement(7387..7405)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(7388..7405)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        complement(7459..7506)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
ORIGIN
        1 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
       61 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
      121 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
      181 acgtcaaagg gcgaaaaacc gtctatcagg gcgctggccc actacgtgaa ccatcaccct
      241 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
      301 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
      361 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
      421 cacccgccgc gcttaatgcg ccgctacagg gcgcgtaaca tgaagtttta aatcaatcta
      481 aagtatatat gtgtaacatt ggtctagtga ttatttgccg actaccttgg tgatctcgcc
      541 tttcacgtag tggacaaatt cttccaactg atctgcgcgc gaggccaaac gatcttcttc
      601 ttgtccaaga taagcctgtc tagcttcaag tatgacgggc tgatactggg ccggcaggcg
      661 ctccattgcc cagtcggcag ctacatcctt cggcgcgatt ttgccggtta ctgcgctgta
      721 ccaaatgcgg gacaacgtaa gcactacatt tcgctcatcg ccagcccagt cgggcggcga
      781 gttccatagc gttaaggttt catttagcgc ctcaaataga tcctgttcag gaaccggatc
      841 aaagagttcc tccgccgctg gacctaccaa ggcaacgcta tgttctcttg cttttgtcag
      901 caagatagcc agatcaatgt cgatcgtggc tggctcgaag atacctgcaa gaatgtcatt
      961 gcgctgccat tctccaaatt gcagttcgcg cttagctgga taacgccacg gaatgatgtc
     1021 gtcgtgcaca acaatggtga cttctacagc gcggagaatc tcgctctctc caggggaagc
     1081 cgaagtttcc aaaaggtcgt tgatcaaagc tcgccgcgtt gtttcatcaa gccttacggt
     1141 caccgtaacc agcaaatcaa tatcactgtg tggcttcagg ccgccatcca ctgcggagcc
     1201 gtacaaatgt acggccagca acgtcggttc gagatggcgc tcgatgacgc caactacctc
     1261 tgatagttga gtcgatactt cggcgatcac cgcttccctc ataatgttta actttgtttt
     1321 agggcgactg ccctgctgcg taacatcgtt gctgctccat aacatcaaac atcgacccac
     1381 ggcgtaacgc gcttgctgct tggatgcccg aggcatagac tgtaccccaa aaaaacagtc
     1441 ataacaagcc atgaaaaccg ccactgcgcc gttaccaccg ctgcgttcgg tcaaggttct
     1501 ggaccagttg cgtgagcgca tacgctactt gcattacagc ttacgaaccg aacaggctta
     1561 tgtccactgg gttcgtgcct tcatccgttt ccacggtgtg cgtcacccgg caaccttggg
     1621 tagcagcgaa gtcgaggcat ttctgtcctg gctggtcaca aaaggatcta ggtgaagatc
     1681 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     1741 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     1801 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     1861 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     1921 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     1981 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     2041 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     2101 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     2161 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     2221 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     2281 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     2341 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     2401 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtgtc
     2461 aagcgcagct gcccgggtct agagtcgacg catgcctcct gacagcggtt ctccttgttc
     2521 tccacggtct taagcttagc acgagcgatc gagagagata gagagcgaga tagagcgaag
     2581 cttgttaatt cgtcgagcgc ttgccatact gaaaaaacct cccacacctc cccctgaacc
     2641 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     2701 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     2761 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatcaag ctcactcctt
     2821 gcgacgagcg cgcaggcgca gagccagata ggccagcacg cggaaggaca caatcaggat
     2881 ggccaggccc acataatcca gtggcaggtc ggcggcactg aaattcagtg tctccaggat
     2941 gacctttccg ctgctggggc aggtcgtgtt gctgctggtg caggagatct cgccgggctc
     3001 cacatcggcc cactgattga tcagcaggcc ctcattggcg tagcgaaacc agctcaggta
     3061 gctcagccac ttcaggtaca cgggcacgct gccggagttc agaaagaatc cgccgaacag
     3121 caggaagggg ataatcacgg ggggtcccac gctcagggcc atgctggtgc tgctggaggc
     3181 gcagctgatc aggtagccga agctggtcga cacgttggcc accagggtca ccagggccag
     3241 gcaattaaag aaatgcagca ctccggcacg caggccaatc atggggtagg caatggcggt
     3301 aaacaccagg ggcacggtca ggaacagcgg cagctcggca atggtctttc ccaggaagta
     3361 ggtatcgcag cggtacaggc ggctgcgggc ctcgcgcatg aacacgggca gctcgctggt
     3421 gaacacgttg atggtggcga acacgttctg gaacgtcatg ttcgtcagaa acagaaaaat
     3481 ggctccattg atgttcatca ctccgacctg cgtcagctgc tgtcccagga agatcaggcc
     3541 gatcagaatg gccaccatcg tggtctggat caggcggacc ttgaccagca ggggctcctt
     3601 cagcacactc agccagctgc gccacagcac ggcgcgaaac tgcataaacc aggtagcctt
     3661 atacgtatag ccgttctcgg gctgctccag gggcttctcc aggttcttcg tggccagcag
     3721 ctgctccatg tcgcgggcca ccttggagat ggcgaagtta tcgcagatct tggcaatgcg
     3781 atcgcggctc tcaatctcgc gtcctggcac cacggccagc acttgcacat agaagtcggc
     3841 ggggttgtaa ttcgtggggc actgggcgcc cacatagctg aagaaatcca cggcctcgct
     3901 tggggttccc aggaaggcca cgcgtccctc ggccatcagc agaatcttat caaacagctc
     3961 gaacagctcc gagctgggct ggtggatcgt cagaatgacc gtctttccct tctggctcag
     4021 cttcttcagc acctgcacca cggagtgggc ggtgaagcta tccaggccgc tggtcggctc
     4081 gtcgcaaatc agcagggggg gatcggtcag ggcctcgctg gcaaaggcca ggcgcttgcg
     4141 ctctccgcca ctcaggccct tcacgcggcc tggcacgcca ataatggtgt gctggcactt
     4201 gctcaggctc agctcctgga tgacctggtc cacgcgggcc acgcgctgac ggtatgtcag
     4261 gtggcggggc atgcggacca tggcctgaaa gatcagatgc tcgcgggctg tcaggctgcc
     4321 gatgaacaga tcgtcctgct gcacgtaggc gcagcgggcc tgcatctcct tggcatccac
     4381 gggctggccg ttcagcaggc gcatgccgct tggggacacc tgaatgccct gggggctgcg
     4441 gaaggccagg gcgttcagca gcgtggtctt tccggcgcca ctacttccca tcacggccag
     4501 cagctcgccg gggtaggcca ctccgcacac attcttcagc aggtgcttgc gtggggctgg
     4561 gatatggcgc tcattgcaga acagtccgcg ggtgcggttc accagctgac gccagccact
     4621 gcctggctgg ttcacggcgc cgaagatatc catgttatgc caggcgtacg tcagattctc
     4681 ggccagctgt ccggatccgc ttccgctatc ctctgggggg ctggggggca gcagggttcc
     4741 gtaattcttg gcctggccaa atccctgatt gatgcacgac tggctggcgg ctccgctatc
     4801 gccgttgttc aggtgctcgg cgcttgggtg cttggagcct ccgcggatca gcagctcctg
     4861 gtcctcctgt cccatttggt ttggtatgcg tcttgtgatt caaagttggc ttattcaaag
     4921 gatattaacg aaggcagcgg cacgtctgct cggctcgaac attcattcac aactgatgct
     4981 ctcagccacc ccgcgccctt ttataccgct gcgctcgatc cccgggcgag ctcgggaggg
     5041 gcatgacgta cctacctcga ctgctactcg acttaaaggc atttcaaggg tttccactgg
     5101 gctcgactta aaggcatttc aagggtttcc actgggctcg acttaaaggc atttcaaggg
     5161 tttccactgg gctcgactta aaggcatttc aagggtttcc actgggctcg acttaaaggc
     5221 atttcaaggg tttccactgg gctcgacagt caggtgctag tcgtacgctc ctgacagcgg
     5281 aacaaacgac ccaacacccg tgcgttttat tctgtctttt tattgccaag cttactcctt
     5341 ggcacgggga cgggtgcttg gacggcgatt tccactatcg gccagcacct ccacgcatcc
     5401 atcggtccac acggcagccg agcgacgggc aatctgggtg cgtcccacgg ttccagctcc
     5461 actacgcaca atggcgtcgc agcgtccctg agcccaggca gcatcatcaa agttgccatc
     5521 caccaggctc tggtagagct ggtccaggcc gatacgcagc atataggcac gcaggcgtgg
     5581 gcttccagcc agctctggat ggcgacgctc aaagtaacgg gtctgctgtt ccatacaggc
     5641 cagccaggga cgccagaaga agatattggc cacctcatac tggctatcgc cgaacatagc
     5701 ctcgctccaa tcaatcacgg ccgtgatacg gccgttatcg gtcagcacat tgttgctgcc
     5761 gaaatcggcg tgcaccaggt gacggacctc tgggcaatcc tcggcccaca gcatcagctc
     5821 gtccagggcc tgggccacgc tggccgacac ggtatcatcc atcacggtct gccagtggta
     5881 gacgtgtggg tcggcaatgg cacagatgaa atcacgccag gtggtatact ggccgatgcc
     5941 ttgtggtccg aatgggccga agccactggt ctggctcagg tcggcagcag caatggcatc
     6001 catagcctca gccactggct ggagcacggc tggcagctcg gtttcgggca ggtcttgcag
     6061 ggtcacgccc tgggcacgac ggctaatgca gtaggtcagg ctctcgctga actcgccgat
     6121 atccagcacc tcggggattg gcagggcagc actggcgaaa tggcgataca cataacggtc
     6181 cttgtagaag ccatcggcac agctgttcac acgcagcaca tatccacggc ctcccacatc
     6241 aaagctgaag gcacggcttt cctcgccctc gctcagctgc atcaggtcgg acacgctatc
     6301 aaacttctca atcaagaact tctcaacgct ggtggccgtc agctcgggct tcttcatttt
     6361 gttttcctcc ttatgttaag cttactcagt tattatatca taaatatctg tgtcaagaat
     6421 aaactcccac atggattcga gtattgagag ttctcttctt gtattcaata attacttctt
     6481 ggcagatttc agtagttgca gttgatttac ttggttgctg gttactttta attgattcac
     6541 tttaacttgc actttactgc agattgttta gcttgttcag ctgcgcttgt ttatttgctt
     6601 agctttcgct tagcgacgtg ttcactttgc ttgtttgaat tgaattgtcg ctccgtagac
     6661 gaagcgcctc tatttatact ccggcgctct tttcgcgaac attcgaggcg cgctctctcg
     6721 aaccaacgag agcagtatgc cgtttactgt gtgacagagt gagagagcat tagtgcagag
     6781 agggagcgac ccaaaaagaa aagagagaat aacgaataac ggccagagaa atttctcgag
     6841 ttttctttct gccaaacaaa tgacctacca caataaccag tttgttttgg gattctagct
     6901 cctgacagcg catcatgatg gaccagatgg gtgaggtgga gtacgcgccc ggggagccca
     6961 agggcacgcc ctggcacccg caccgcggct tcgagacagt gacctacatc gtcgaccaat
     7021 tcctccctgg gatccggaga gagaccctat gcggcatcag agcagattgt actgagagtg
     7081 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgt
     7141 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     7201 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
     7261 ttttcccagt cacgacgttg taaaacgacg gccagtgaat tgtaatacga ctcactatag
     7321 ggcgaattcg agctcggtac ccggggatcc tctagagtcg acctgcaggc atgcaagctt
     7381 gagtattcta tagtgtcacc taaatagctt ggcgtaatca tggtcatgat ctgcctcctg
     7441 ggtttagttc ctcaccttgt cgtattatac tatgccgata tactatgccg atgattaatt
     7501 gtcaacaggg gtctctcgct gatccgtta
//
