LOCUS       sequence-108846-        4324 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses drosophila IMD tagged with V5 and 6xHis epitopes, under 
            control of metallothionein promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4324)
  TITLE     pMT-IMD-V5-His
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4324)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pMT-IMD-V5-His
FEATURES             Location/Qualifiers
     source          1..4324
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        401..827
                     /label=MT promoter
                     /note="Drosophila metallothionein promoter"
     CDS             1716..1757
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1767..1784
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1807..1824)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1826..1960
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1850..1869)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1904..1923
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2104..2120)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2104..2120)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2117..2139)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2128..2144
                     /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(2152..2158,2159..2176,2177..2182))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2197..2218
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2335..2352)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2506..3094)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2586..2605)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3265..4056,4057..4125))
                     /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     3888..3907
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4126..4230)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4298..4316
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt aattcgttgc aggacaggat
      421 gtggtgcccg atgtgactag ctctttgctg caggccgtcc tatcctctgg ttccgataag
      481 agacccagaa ctccggcccc ccaccgccca ccgccacccc catacatatg tggtacgcaa
      541 gtaagagtgc ctgcgcatgc cccatgtgcc ccaccaagag ctttgcatcc catacaagtc
      601 cccaaagtgg agaaccgaac caattcttcg cgggcagaac aaaagcttct gcacacgtct
      661 ccactcgaat ttggagccgg ccggcgtgtg caaaagaggt gaatcgaacg aaagacccgt
      721 gtgtaaagcc gcgtttccaa aatgtataaa accgagagca tctggccaat gtgcatcagt
      781 tgtggtcagc agcaaaatca agtgaatcat ctcagtgcaa ctaaaggggg gatctagatc
      841 ggggtaccta ctagtccagt gtggtggaat tcatgtcaaa gctcaggaac ctgttgccca
      901 caatctttgg cgggaaggag gcacagaatc cgacaccagt cgagggacgc ctggaaaagg
      961 acgcagctcc cgtggacgac aacgaaccag ataacaacaa cagcggagcc atggcgctgc
     1021 cctccaccgc tggcacgccc acagcctcct cggatctgac cgaatccgtg ctgcgcgagc
     1081 tcagcgaccc aaactacaat tcaatggatg tggtgcattc cgccaatatt ccgggcactc
     1141 tcagtaacgt ccagacaaac aacaccatga acgtacacag cgcccagcaa caggtggtca
     1201 tgaacttctc gaatgccaat aatctgcact tcggctccgt ctacaacttc aaccaaaact
     1261 tgagcgcctg cagctcgcga aagggtagca ccagtaccgc agaggagtcg gtcgcctctc
     1321 cagacggtaa gccgcgggca agtgcgacgc gcaaaacggt cagcattgtg gccatgatgc
     1381 agtcacaaga ggaaccggat gtgcgcctgc tcgatgtggt ttccacgcac ttgggcgagg
     1441 gctggaagca ggtgatgcgg gatcttggca tgtcggaagg acagatcgac caggccataa
     1501 tcgattacca aatgcatggc aatattagag aggtgatata ccagctgttg ctccagtgga
     1561 ttcgaagctc ggcggacggt gtggccaccg ttggacgact cactacactg ctgtgggagt
     1621 cccagcatcg cgactgcgtg cagcgcatga aactggtttg gaaggcactg gagaagcgca
     1681 agacaaacag cctcgagtct agagggccct tcgaaggtaa gcctatccct aaccctctcc
     1741 tcggtctcga ttctacgcgt accggtcatc atcaccatca ccattgagtt taaacccgct
     1801 gatcagcctc gactgtgcct tctaagatcc agacatgata agatacattg atgagtttgg
     1861 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     1921 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     1981 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     2041 caaatgtggt atggctgatt atgatcagtc gacctgcagg catgcaagct tggcgtaatc
     2101 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     2161 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     2221 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     2281 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     2341 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     2401 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     2461 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     2521 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     2581 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     2641 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     2701 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     2761 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     2821 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     2881 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     2941 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     3001 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     3061 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     3121 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     3181 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     3241 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     3301 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     3361 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     3421 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     3481 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     3541 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     3601 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     3661 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     3721 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     3781 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     3841 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     3901 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     3961 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     4021 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     4081 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     4141 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     4201 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     4261 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     4321 tcgt
//
