LOCUS       sequence-329257-        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"
     promoter        24..71
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        125..143
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     125..142
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     misc_feature    150..206
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(208..227)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(209..227)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(234..251)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(234..250)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(243..265)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(459..478)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     protein_bind    535..604
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     promoter        632..1083
                     /label=hsp70 promoter
                     /note="Drosophila melanogaster hsp70Bb promoter"
     primer_bind     1031..1052
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     CDS             1173..2198
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    4723..4857
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4747..4766)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4801..4820
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(5172..5760)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5252..5271)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             6228..7019
                     /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      7074..7502
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7161..7180)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7371..7392
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 taacggatca gcgagagacc cctgttgaca attaatcatc ggcatagtat atcggcatag
       61 tataatacga caaggtgagg aactaaaccc aggaggcaga tcatgaccat gattacgcca
      121 agctatttag gtgacactat agaatactca agcttgcatg cctgcaggtc gactctagag
      181 gatccccggg taccgagctc gaattcgccc tatagtgagt cgtattacaa ttcactggcc
      241 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
      301 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
      361 caacagttgc gcagcctgaa tggcgaatga cgcctgatgc ggtattttct ccttacgcat
      421 ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgca
      481 tagggtctct ctccggatcc cagggaggaa ttggtcgacg atgtaggtca ctgtctcgaa
      541 gccgcggtgc gggtgccagg gcgtgccctt gggctccccg ggcgcgtact ccacctcacc
      601 catctggtcc atcatgatgc gctgtcagga gctagaatcc caaaacaaac tggttattgt
      661 ggtaggtcat ttgtttggca gaaagaaaac tcgagaaatt tctctggccg ttattcgtta
      721 ttctctcttt tctttttggg tcgctccctc tctgcactaa tgctctctca ctctgtcaca
      781 cagtaaacgg catactgctc tcgttggttc gagagagcgc gcctcgaatg ttcgcgaaaa
      841 gagcgccgga gtataaatag aggcgcttcg tctacggagc gacaattcaa ttcaaacaag
      901 caaagtgaac acgtcgctaa gcgaaagcta agcaaataaa caagcgcagc tgaacaagct
      961 aaacaatctg cagtaaagtg caagttaaag tgaatcaatt aaaagtaacc agcaaccaag
     1021 taaatcaact gcaactactg aaatctgcca agaagtaatt attgaataca agaagagaac
     1081 tctcaatact cgaatccatg tgggagttta ttcttgacac agatatttat gatataataa
     1141 ctgagtaagc ttaacataag gaggaaaaca aaatgaagaa gcccgagctg acggccacca
     1201 gcgttgagaa gttcttgatt gagaagtttg atagcgtgtc cgacctgatg cagctgagcg
     1261 agggcgagga aagccgtgcc ttcagctttg atgtgggagg ccgtggatat gtgctgcgtg
     1321 tgaacagctg tgccgatggc ttctacaagg accgttatgt gtatcgccat ttcgccagtg
     1381 ctgccctgcc aatccccgag gtgctggata tcggcgagtt cagcgagagc ctgacctact
     1441 gcattagccg tcgtgcccag ggcgtgaccc tgcaagacct gcccgaaacc gagctgccag
     1501 ccgtgctcca gccagtggct gaggctatgg atgccattgc tgctgccgac ctgagccaga
     1561 ccagtggctt cggcccattc ggaccacaag gcatcggcca gtataccacc tggcgtgatt
     1621 tcatctgtgc cattgccgac ccacacgtct accactggca gaccgtgatg gatgataccg
     1681 tgtcggccag cgtggcccag gccctggacg agctgatgct gtgggccgag gattgcccag
     1741 aggtccgtca cctggtgcac gccgatttcg gcagcaacaa tgtgctgacc gataacggcc
     1801 gtatcacggc cgtgattgat tggagcgagg ctatgttcgg cgatagccag tatgaggtgg
     1861 ccaatatctt cttctggcgt ccctggctgg cctgtatgga acagcagacc cgttactttg
     1921 agcgtcgcca tccagagctg gctggaagcc cacgcctgcg tgcctatatg ctgcgtatcg
     1981 gcctggacca gctctaccag agcctggtgg atggcaactt tgatgatgct gcctgggctc
     2041 agggacgctg cgacgccatt gtgcgtagtg gagctggaac cgtgggacgc acccagattg
     2101 cccgtcgctc ggctgccgtg tggaccgatg gatgcgtgga ggtgctggcc gatagtggaa
     2161 atcgccgtcc aagcacccgt ccccgtgcca aggagtaagc ttggcaataa aaagacagaa
     2221 taaaacgcac gggtgttggg tcgtttgttc cgctgtcagg agcgtacgac tagcacctga
     2281 ctgtcgagcc cagtggaaac ccttgaaatg cctttaagtc gagcccagtg gaaacccttg
     2341 aaatgccttt aagtcgagcc cagtggaaac ccttgaaatg cctttaagtc gagcccagtg
     2401 gaaacccttg aaatgccttt aagtcgagcc cagtggaaac ccttgaaatg cctttaagtc
     2461 gagtagcagt cgaggtaggt acgtcatgcc cctcccgagc tcgcccgggg atcgagcgca
     2521 gcggtataaa agggcgcggg gtggctgaga gcatcagttg tgaatgaatg ttcgagccga
     2581 gcagacgtgc cgctgccttc gttaatatcc tttgaataag ccaactttga atcacaagac
     2641 gcataccaaa ccaaatggga caggaggacc aggagctgct gatccgcgga ggctccaagc
     2701 acccaagcgc cgagcacctg aacaacggcg atagcggagc cgccagccag tcgtgcatca
     2761 atcagggatt tggccaggcc aagaattacg gaaccctgct gccccccagc cccccagagg
     2821 atagcggaag cggatccgga cagctggccg agaatctgac gtacgcctgg cataacatgg
     2881 atatcttcgg cgccgtgaac cagccaggca gtggctggcg tcagctggtg aaccgcaccc
     2941 gcggactgtt ctgcaatgag cgccatatcc cagccccacg caagcacctg ctgaagaatg
     3001 tgtgcggagt ggcctacccc ggcgagctgc tggccgtgat gggaagtagt ggcgccggaa
     3061 agaccacgct gctgaacgcc ctggccttcc gcagccccca gggcattcag gtgtccccaa
     3121 gcggcatgcg cctgctgaac ggccagcccg tggatgccaa ggagatgcag gcccgctgcg
     3181 cctacgtgca gcaggacgat ctgttcatcg gcagcctgac agcccgcgag catctgatct
     3241 ttcaggccat ggtccgcatg ccccgccacc tgacataccg tcagcgcgtg gcccgcgtgg
     3301 accaggtcat ccaggagctg agcctgagca agtgccagca caccattatt ggcgtgccag
     3361 gccgcgtgaa gggcctgagt ggcggagagc gcaagcgcct ggcctttgcc agcgaggccc
     3421 tgaccgatcc ccccctgctg atttgcgacg agccgaccag cggcctggat agcttcaccg
     3481 cccactccgt ggtgcaggtg ctgaagaagc tgagccagaa gggaaagacg gtcattctga
     3541 cgatccacca gcccagctcg gagctgttcg agctgtttga taagattctg ctgatggccg
     3601 agggacgcgt ggccttcctg ggaaccccaa gcgaggccgt ggatttcttc agctatgtgg
     3661 gcgcccagtg ccccacgaat tacaaccccg ccgacttcta tgtgcaagtg ctggccgtgg
     3721 tgccaggacg cgagattgag agccgcgatc gcattgccaa gatctgcgat aacttcgcca
     3781 tctccaaggt ggcccgcgac atggagcagc tgctggccac gaagaacctg gagaagcccc
     3841 tggagcagcc cgagaacggc tatacgtata aggctacctg gtttatgcag tttcgcgccg
     3901 tgctgtggcg cagctggctg agtgtgctga aggagcccct gctggtcaag gtccgcctga
     3961 tccagaccac gatggtggcc attctgatcg gcctgatctt cctgggacag cagctgacgc
     4021 aggtcggagt gatgaacatc aatggagcca tttttctgtt tctgacgaac atgacgttcc
     4081 agaacgtgtt cgccaccatc aacgtgttca ccagcgagct gcccgtgttc atgcgcgagg
     4141 cccgcagccg cctgtaccgc tgcgatacct acttcctggg aaagaccatt gccgagctgc
     4201 cgctgttcct gaccgtgccc ctggtgttta ccgccattgc ctaccccatg attggcctgc
     4261 gtgccggagt gctgcatttc tttaattgcc tggccctggt gaccctggtg gccaacgtgt
     4321 cgaccagctt cggctacctg atcagctgcg cctccagcag caccagcatg gccctgagcg
     4381 tgggaccccc cgtgattatc cccttcctgc tgttcggcgg attctttctg aactccggca
     4441 gcgtgcccgt gtacctgaag tggctgagct acctgagctg gtttcgctac gccaatgagg
     4501 gcctgctgat caatcagtgg gccgatgtgg agcccggcga gatctcctgc accagcagca
     4561 acacgacctg ccccagcagc ggaaaggtca tcctggagac actgaatttc agtgccgccg
     4621 acctgccact ggattatgtg ggcctggcca tcctgattgt gtccttccgc gtgctggcct
     4681 atctggctct gcgcctgcgc gctcgtcgca aggagtgagc ttgatccaga catgataaga
     4741 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     4801 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     4861 aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttca
     4921 gtatggcaag cgctcgacga attaacaagc ttcgctctat ctcgctctct atctctctcg
     4981 atcgctcgtg ctaagcttaa gaccgtggag aacaaggaga accgctgtca ggaggcatgc
     5041 gtcgactcta gacccgggca gctgcgcttg acacggttat ccacagaatc aggggataac
     5101 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     5161 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     5221 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     5281 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     5341 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     5401 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     5461 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     5521 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     5581 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     5641 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     5701 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     5761 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     5821 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt gtgaccagcc
     5881 aggacagaaa tgcctcgact tcgctgctac ccaaggttgc cgggtgacgc acaccgtgga
     5941 aacggatgaa ggcacgaacc cagtggacat aagcctgttc ggttcgtaag ctgtaatgca
     6001 agtagcgtat gcgctcacgc aactggtcca gaaccttgac cgaacgcagc ggtggtaacg
     6061 gcgcagtggc ggttttcatg gcttgttatg actgtttttt tggggtacag tctatgcctc
     6121 gggcatccaa gcagcaagcg cgttacgccg tgggtcgatg tttgatgtta tggagcagca
     6181 acgatgttac gcagcagggc agtcgcccta aaacaaagtt aaacattatg agggaagcgg
     6241 tgatcgccga agtatcgact caactatcag aggtagttgg cgtcatcgag cgccatctcg
     6301 aaccgacgtt gctggccgta catttgtacg gctccgcagt ggatggcggc ctgaagccac
     6361 acagtgatat tgatttgctg gttacggtga ccgtaaggct tgatgaaaca acgcggcgag
     6421 ctttgatcaa cgaccttttg gaaacttcgg cttcccctgg agagagcgag attctccgcg
     6481 ctgtagaagt caccattgtt gtgcacgacg acatcattcc gtggcgttat ccagctaagc
     6541 gcgaactgca atttggagaa tggcagcgca atgacattct tgcaggtatc ttcgagccag
     6601 ccacgatcga cattgatctg gctatcttgc tgacaaaagc aagagaacat agcgttgcct
     6661 tggtaggtcc agcggcggag gaactctttg atccggttcc tgaacaggat ctatttgagg
     6721 cgctaaatga aaccttaacg ctatggaact cgccgcccga ctgggctggc gatgagcgaa
     6781 atgtagtgct tacgttgtcc cgcatttggt acagcgcagt aaccggcaaa atcgcgccga
     6841 aggatgtagc tgccgactgg gcaatggagc gcctgccggc ccagtatcag cccgtcatac
     6901 ttgaagctag acaggcttat cttggacaag aagaagatcg tttggcctcg cgcgcagatc
     6961 agttggaaga atttgtccac tacgtgaaag gcgagatcac caaggtagtc ggcaaataat
     7021 cactagacca atgttacaca tatatacttt agattgattt aaaacttcat gttacgcgcc
     7081 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     7141 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     7201 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     7261 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccagcgcc
     7321 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     7381 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     7441 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     7501 ttttaacaaa atattaacgc ttacaattt
//
