LOCUS       sequence-374439-        5821 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Plasmid for the protein expression of GST-mCherry-ATG14 in a 
            bacterial and insect expression system. Internal_Reference: SMC1322.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5821)
  TITLE     ASAP - PI3KC3-C1 related projects
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5821)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pGB-05-06-GST-mCherry-ATG14
FEATURES             Location/Qualifiers
     source          1..5821
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     mobile_element  17..240
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(307..840)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(1029..1034,1035..1051,1052..1057))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(1128..1161)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        1250..1341
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     1266..1283
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             1397..2050
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2008..2030
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2057..2080
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2081..2785
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(2223..2241)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2491..2510
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             4333..4350
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4494..4628
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4531..4550)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4585..4604
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  4760..4925
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      5115..5703
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5604..5623
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gccctgcgta agcgggtgtg ggcggacaat aaagtcttaa actgaacaaa atagatctaa
       61 actatgacaa taaagtctta aactagacag aatagttgta aactgaaatc agtccagtta
      121 tgctgtgaaa aagcatactg gacttttgtt atggctaaag caaactcttc attttctgaa
      181 gtgcaaattg cccgtcgtat taaagagggg cgtggccaag ggcatgtaaa gactatattc
      241 gcggcgttgt gacaatttac cgaacaactc cgcggccggg aagccgatct cggcttgaac
      301 gaattgttag gtggcggtac ttgggtcgat atcaaagtgc atcacttctt cccgtatgcc
      361 caactttgta tagagagcca ctgcgggatc gtcaccgtaa tctgcttgca cgtagatcac
      421 ataagcacca agcgcgttgg cctcatgctt gaggagattg atgagcgcgg tggcaatgcc
      481 ctgcctccgg tgctcgccgg agactgcgag atcatagata tagatctcac tacgcggctg
      541 ctcaaacttg ggcagaacgt aagccgcgag agcgccaaca accgcttctt ggtcgaaggc
      601 agcaagcgcg atgaatgtct tactacggag caagttcccg aggtaatcgg agtccggctg
      661 atgttgggag taggtggcta cgtctccgaa ctcacgaccg aaaagatcaa gagcagcccg
      721 catggatttg acttggtcag ggccgagcct acatgtgcga atgatgccca tacttgagcc
      781 acctaacttt gttttagggc gactgccctg ctgcgtaaca tcgttgctgc tgcgtaacat
      841 cgttgctgct ccataacatc aaacatcgac ccacggcgta acgcgcttgc tgcttggatg
      901 cccgaggcat agactgtaca aaaaaacagt cataacaagc catgaaaacc gccactgcgc
      961 cgttaccacc gctgcgttcg gtcaaggttc tggaccagtt gcgtgagcgc atacgctact
     1021 tgcattacag tttacgaacc gaacaggctt atgtcaactg ggttcgtgcc ttcatccgtt
     1081 tccacggtgt gcgtcacccg gcaaccttgg gcagcagcga agtcgccata acttcgtata
     1141 gcatacatta tacgaagtta tctgtaacta taacggtcct aaggtagcga aggtctcact
     1201 gcgtttaaac actagtatcg attcgcgacc tactccggaa tattaataga tcatggagat
     1261 aattaaaatg ataaccatct cgcaaataaa taagtatttt actgttttcg taacagtttt
     1321 gtaataaaaa aacctataaa tattccggat tattcatacc gtcccaccat cgggcgacgg
     1381 atcccggtcc gaaaccatgt cccctatact aggttattgg aaaattaagg gccttgtgca
     1441 acccactcga cttcttttgg aatatcttga agaaaaatat gaagagcatt tgtatgagcg
     1501 cgatgaaggt gataaatggc gaaacaaaaa gtttgaattg ggtttggagt ttcccaatct
     1561 tccttattat attgatggtg atgttaaatt aacacagtct atggccatca tacgttatat
     1621 agctgacaag cacaacatgt tgggtggttg tccaaaagag cgtgcagaga tttcaatgct
     1681 tgaaggagcg gttttggata ttagatacgg tgtttcgaga attgcatata gtaaagactt
     1741 tgaaactctc aaagttgatt ttcttagcaa gctacctgaa atgctgaaaa tgttcgaaga
     1801 tcgtttatgt cataaaacat atttaaatgg tgatcatgta acccatcctg acttcatgtt
     1861 gtatgacgct cttgatgttg ttttatacat ggacccaatg tgcctggatg cgttcccaaa
     1921 attagtttgt tttaaaaaac gtattgaagc tatcccacaa attgataagt acttgaaatc
     1981 cagcaagtat atagcatggc ctttgcaggg ctggcaagcc acgtttggtg gtggcgacca
     2041 tcctccaaaa tcggatctgg aagttctgtt ccaggggccc gtgagcaagg gcgaggagga
     2101 taacatggcc atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa
     2161 cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac
     2221 cgccaagctg aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc
     2281 tcagttcatg tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt
     2341 gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg
     2401 cgtggtgacc gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa
     2461 gctgcgcggc accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg
     2521 ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca
     2581 gaggctgaag ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc
     2641 caagaagccc gtgcagctgc ccggcgccta caacgtcaac atcaagttgg acatcacctc
     2701 ccacaacgag gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac
     2761 cggcggcatg gacgagctgt acaagggttc cggctctggt agcatggcat cgccaagcgg
     2821 aaaaggagca agagccctgg aagcacccgg ctgcggacca agaccactcg cccgtgacct
     2881 cgttgatagc gtggacgacg ctgagggtct gtacgtggcc gtcgaacgtt gccctctgtg
     2941 caacaccact cgccgtaggc tgacctgcgc taagtgcgtg cagagcggtg acttcgtcta
     3001 cttcgacgga cgcgaccgcg agcgtttcat cgacaagaag gaaaggctgt ccagactgaa
     3061 gagcaagcag gaggagttcc agaaggaggt cctgaaggct atggagggca agtggatcac
     3121 tgaccagctg cgctggaaga tcatgtcttg caagatgcgt atcgagcagc tgaagcagac
     3181 catctgcaag ggcaacgagg aaatggagaa gaactctgaa ggactgctga agactaagga
     3241 aaagaaccag aagctgtact cacgcgctca gcgtcaccag gagaagaagg aaaagatcca
     3301 gaggcacaac agaaagctgg gtgacctggt ggagaagaag accatcgacc tgcgcagcca
     3361 ctacgaacgt ctggccaacc tgagacgctc ccacatcctg gagctgacca gcgtcatctt
     3421 ccccatcgag gaagtgaaga ctggcgtcag ggacccagct gacgtgtcca gcgagagcga
     3481 ctctgccatg acctcttcaa ctgtctccaa gctggctgaa gcccgtagga ccacttacct
     3541 gagcggacgc tgggtgtgcg acgaccacaa cggagacacc tcaatctcca tcactggtcc
     3601 ctggatctcc ctgccaaaca acggagacta ctctgcttac tactcatggg tcgaggaaaa
     3661 gaagaccact cagggtcctg acatggagca gtctaacccc gcttacacca tctcagctgc
     3721 cctgtgctac gccactcagc tggtgaacat cctgtctcac atcctggacg tcaacctgcc
     3781 aaagaagctg tgcaactccg agttctgcgg cgaaaacctg agcaagcaga agttcacccg
     3841 cgctgtgaag aagctgaacg ccaacatcct gtacctgtgc ttctcacagc acgtcaacct
     3901 ggaccagctg caacctctgc acactctgcg taacctgatg tacctggtgt ctccatccag
     3961 cgagcacctg ggaaggtcag gtccattcga agtcagagct gacctggagg aaagcatgga
     4021 gttcgtggac cctggcgtcg ccggagagtc tgacgaatca ggtgacgaga gggtgtccga
     4081 cgaggaaacc gacctgggca ctgactggga aaacctgcca agccctagat tctgcgacat
     4141 ccccagccag tctgtggaag tctcacaatc tcagagcacc caggcctccc ctcccatcgc
     4201 ttcttcatcc gccggtggca tgatctcgtc agccgcagca tccgtcactt cgtggttcaa
     4261 ggcatacact ggacataggt aacgccatta acctgatgtt ctggggaata taagcttgcg
     4321 gccgcactcg agcaccacca ccaccaccac tgacatgcgg taccaagctt gtcgagaagt
     4381 actagaggat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     4441 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     4501 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     4561 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     4621 tctggatctg atcactgctt gagcctagaa gatccggctg ctaacaaagc ccgaaaggaa
     4681 gctgagttgg ctgctgccac cgctgagcaa taactatcat aacccctagg gtatacccat
     4741 ctaattggac tatgagacca accagataag tgaaatctag ttccaaacta ttttgtcatt
     4801 tttaattttc gtattagctt acgacgctac acccagttcc catctatttt gtcactcttc
     4861 cctaaataat ccttaaaaac tccatttcca cccctcccag ttcccaacta ttttgtccgc
     4921 ccacaaccgg ttgacttggg tcaactgtca gaccaagttt actcatatat actttagatt
     4981 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     5041 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     5101 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     5161 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     5221 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     5281 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     5341 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     5401 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     5461 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     5521 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     5581 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     5641 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     5701 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     5761 atgttctttc ctgcgttatc ccctgattga cttgggtcgc tcttcctgtg gatgcgcaga
     5821 t
//
