LOCUS       sequence-345195-        5459 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Drosophila Alpha Tubulin promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5459)
  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 5459)
  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: pDmAlfaTub84b
FEATURES             Location/Qualifiers
     source          1..5459
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     163..182
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     416..433
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    550..571
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(586..591,592..609,610..616)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    624..640
                     /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)."
     primer_bind     629..651
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     648..664
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     648..664
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        682..700
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     682..699
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      3238..3247
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3242..3251
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3281..3300)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3282..3300)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3307..3324)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3307..3323)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3316..3338)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3464..3919
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3551..3570)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3761..3782
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3997..4101
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4102..4170,4171..4962)
                     /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     complement(4320..4339)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(5133..5459,1..262)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
       61 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
      121 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
      181 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
      241 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
      301 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
      361 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
      421 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
      481 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
      541 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct
      601 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta
      661 tgaccatgat tacgccaagc tatttaggtg acactataga atactcaagc tatgcatcca
      721 acgcgttggg agctctccca tatggtcgac ctgcaggcgg ccgcgaattc actagtgatt
      781 gggcgatgag tggtctcggg agaaaacgaa cgtattgctt agggacacag ctcacattaa
      841 taagacgcca gtcctcgctg tcgcagaaga acttcctcat agcccgcagc ttggcgacgg
      901 tagcgttgga agctcgatgc tcgaacgtgt gaaatgtgtt gcctgccaat tcagaagcca
      961 ggtgaaattg gtaggagctc aatgcagtac taacccgact taccgatgtc gacgaagagg
     1021 tccaggtccg aggatcgatt tccaatgcct gtgatgcggg acccgaactt gtagactcgc
     1081 agtggttgtt tgggaaagcc cggcgacaaa agcttgcaca ggtcctgttc gataacgtcg
     1141 tactcgggaa tcacctttaa cttgtccgct attgggtatt tgtccttaaa cacattcgta
     1201 atctctccca caatcattgc cctggcctct gcaatgtaaa aagtatttat aaaatatcgc
     1261 tggctctaaa gaaaacatat taatcaaagt attgaggctt tatcaatcta acatttacaa
     1321 gtagtcagac cacaattttt agactgcact tctgcgtaag taaatgaaat gaaatggtga
     1381 cctactcttg ggtagccgta ctgttatatg ctgacgagcc ttcacctggg cctgcttttt
     1441 ctcctgtttc gtctgatgcg tcggatgcgg ctggactggc tgattgttgg gattgggttt
     1501 tggatgctgc tccgaaaggt gggccacagc ttttgcgatg gtggtgccca ctatgcggtt
     1561 gctgcaggaa ctacagctga atgttcgctt tattgtctgc agctcctggg gctggacgaa
     1621 ggccacgccc tcacccggtg cggccatctc ctgcggcaaa acgtatgtgg gattcctgtc
     1681 caagtagctc tcgatgccca ccataatgcg ctccagcagg ttggccaggt gcgggtctag
     1741 ctgcggcgga ttcgtcgtct ggttcactga gtttagggtc tgcagaaaga agtgtgcctc
     1801 catggtaagc ggattcatct gcacgatcag cgggctttag tctaaaaatt cgaatgatac
     1861 aaactcgtac tcacgtaaag cagtttgttt ggcatcatct cggccatcgt cttattcttc
     1921 ttcgtggcct ttgttaccgg agcggctatg ttgggcgcgg tggagcccgg aagctgaaag
     1981 gttaatcgaa tgcgttaact tttctgcaac tcgaaagttt gccgcctttg ttcgactgcc
     2041 aataactgtt gattcgaaaa ttcgaatcga agcggttgaa tttcgtaggg tggccaacta
     2101 caccaaagtt cgccggcggt gtatgcatgt gagtgtgagt gtgacagtgg gtgtacgtgt
     2161 gctttaagcg acacaatcgc accgcgatgt acatattttt gtagaaatgt gtgtgcattt
     2221 ttcgtccgga tcacggctcc ataaaataaa tcatatcaca gggctcaggg atttgtgagt
     2281 agaagttccc gccacttaaa cgtcacttgt gaaatcgcca tcgacggtca acttttactc
     2341 acactggtct tccgcggacg gacagtctca aagtactgcc tttctgcgtt ggaaaacatc
     2401 gcctttttcg tccaaaagga gtccccaggt tcgatccgca tggcgttgtg cgtgcgtgcc
     2461 tttcttttca aatgattacg gctattaact tgggggcgtt aagttggaaa cacgtaaatt
     2521 gcagactgcg attagagtga ccatgagtag gagttcaaaa tctcctgaca tcattttctt
     2581 aaaacctgct ttgtttttta catttctatt taatataact cctatttgaa taaaaaaaca
     2641 aaacaagttt agatgttaag atattaacta catcctttgc tccaaaggga gaggggaagt
     2701 tatggagtta attaatttgc tgttggaaat caatatggag tcagaaatat aatgatttac
     2761 taaaccttat tgaatcggta acgatgcgaa tttatattaa aatagctttt atgaaacatt
     2821 caacaaaaat attattaatg ttggcccact ttagcaaccg gttaggtcta ccggttgggc
     2881 aagcaaagat tcacgccctg gttcgagtcc caactagtcc tgcaaaatac cgcagcaagt
     2941 tttagataga ccaagtgcca ttacctctcc cacttcagtt atcggttatg cggcgtttaa
     3001 gtcgacagct tgccgtctat agctccggtg cctatataaa gcagcccgct ttccacattt
     3061 catattcgtt ttacgtttgt caagcctcat agccggcagt tcgaacgtat acgctctctg
     3121 agtcagacct cgaaatcgta gctctacaca attctgtgaa ttttccttgt cgcgtgtgaa
     3181 acacttccaa taaaaactca aatgcgagac cactcatcgc ccaatcgaat tcccgcggcc
     3241 gccatggcgg ccgggagcat gcgacgtcgg gcccaattcg ccctatagtg agtcgtatta
     3301 caattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
     3361 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
     3421 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggacgcgcc ctgtagcggc
     3481 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     3541 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     3601 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     3661 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccagcgcc ctgatagacg
     3721 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     3781 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     3841 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     3901 atattaacgc ttacaatttc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     3961 cacaccgcat caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     4021 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     4081 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     4141 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     4201 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     4261 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     4321 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     4381 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     4441 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     4501 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     4561 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     4621 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     4681 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     4741 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     4801 ggagccggtg agcgtgggtc gcgcggtatc attgcagcac tggggccaga tggtaagccc
     4861 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     4921 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     4981 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     5041 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     5101 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     5161 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     5221 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     5281 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     5341 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     5401 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacgggggg
//
