LOCUS       sequence-323310-        4084 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  hygromycin B resistance marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4084)
  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 4084)
  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: pHPH
FEATURES             Location/Qualifiers
     source          1..4084
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     17..36
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        17..35
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      66..75
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      70..79
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(117..1142)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     promoter        complement(1242..1260)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1243..1260)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1278..1294)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1278..1294)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1291..1313)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1302..1318
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1326..1332,1333..1350,1351..1356))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1371..1392
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1509..1526)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1680..2268)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1760..1779)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2439..3230,3231..3299))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3062..3081
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3300..3404)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3482..3937)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3619..3640)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3831..3850
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     join(4063..4084,1)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4077..4084,1..10)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     join(4078..4084,1..10)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gacggccagt gaattgtaat acgactcact atagggcgaa ttgggcccga cgtcgcatgc
       61 tcccggccgc catggcggcc gcgggaattc gattgggcga tgagtggtct cgaagcttac
      121 tccttggcac ggggacgggt gcttggacgg cgatttccac tatcggccag cacctccacg
      181 catccatcgg tccacacggc agccgagcga cgggcaatct gggtgcgtcc cacggttcca
      241 gctccactac gcacaatggc gtcgcagcgt ccctgagccc aggcagcatc atcaaagttg
      301 ccatccacca ggctctggta gagctggtcc aggccgatac gcagcatata ggcacgcagg
      361 cgtgggcttc cagccagctc tggatggcga cgctcaaagt aacgggtctg ctgttccata
      421 caggccagcc agggacgcca gaagaagata ttggccacct catactggct atcgccgaac
      481 atagcctcgc tccaatcaat cacggccgtg atacggccgt tatcggtcag cacattgttg
      541 ctgccgaaat cggcgtgcac caggtgacgg acctctgggc aatcctcggc ccacagcatc
      601 agctcgtcca gggcctgggc cacgctggcc gacacggtat catccatcac ggtctgccag
      661 tggtagacgt gtgggtcggc aatggcacag atgaaatcac gccaggtggt atactggccg
      721 atgccttgtg gtccgaatgg gccgaagcca ctggtctggc tcaggtcggc agcagcaatg
      781 gcatccatag cctcagccac tggctggagc acggctggca gctcggtttc gggcaggtct
      841 tgcagggtca cgccctgggc acgacggcta atgcagtagg tcaggctctc gctgaactcg
      901 ccgatatcca gcacctcggg gattggcagg gcagcactgg cgaaatggcg atacacataa
      961 cggtccttgt agaagccatc ggcacagctg ttcacacgca gcacatatcc acggcctccc
     1021 acatcaaagc tgaaggcacg gctttcctcg ccctcgctca gctgcatcag gtcggacacg
     1081 ctatcaaact tctcaatcaa gaacttctca acgctggtgg ccgtcagctc gggcttcttc
     1141 attcgagacc actcatcgcc caatcactag tgaattcgcg gccgcctgca ggtcgaccat
     1201 atgggagagc tcccaacgcg ttggatgcat agcttgagta ttctatagtg tcacctaaat
     1261 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     1321 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     1381 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     1441 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     1501 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     1561 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     1621 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     1681 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     1741 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     1801 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     1861 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     1921 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     1981 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     2041 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     2101 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     2161 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     2221 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     2281 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     2341 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     2401 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     2461 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     2521 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcgc
     2581 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     2641 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     2701 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     2761 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     2821 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     2881 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     2941 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     3001 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     3061 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     3121 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     3181 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     3241 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     3301 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     3361 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     3421 gtgccacctg atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     3481 gaaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     3541 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     3601 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     3661 aacgtcaaag ggcgaaaaac cgtctatcag ggcgctggcc cactacgtga accatcaccc
     3721 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     3781 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     3841 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     3901 acacccgccg cgcttaatgc gccgctacag ggcgcgtcca ttcgccattc aggctgcgca
     3961 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
     4021 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
     4081 aaac
//
