LOCUS       sequence-477836-        5806 bp    DNA     circular SYN 07-JUL-2025
DEFINITION  Template for in vitro transcription of PMEL (gp100), including 
            native signal peptide, flanked by the human haemoglobin beta 5' and 
            3' UTRs..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5806)
  AUTHORS   Smart AD, Hughes ME, Ruiz Velasco AD, Hori N, Stolnik S, Jopling CL
  TITLE     A proximity labelling-based approach to directly detect mRNA 
            delivery to specific subcellular locations
  JOURNAL   Molecular Therapy Nucleic Acids, 102602. doi: 
            10.1016/j.omtn.2025.102602
REFERENCE   2  (bases 1 to 5806)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 7, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Molecular 
            Therapy Nucleic Acids, 102602. doi: 10.1016/j.omtn.2025.102602"
COMMENT     Sequence Label: pPMEL-hbb
FEATURES             Location/Qualifiers
     source          1..5806
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        48..244
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      95..230
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     268..287
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        268..286
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2523..2644
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2534..2553)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2588..2607
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2966..2986
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     complement(3138..3155)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3309..3897)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3389..3408)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4068..4859,4860..4928))
                     /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     4691..4710
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4929..5033)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5060..5515
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5147..5166)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5357..5378
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5646..5694
                     /note="synthetic polyadenylation signal"
     misc_feature    5708..5799
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5748..5767
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcttacgcgt gctagcccgg gctcgagatc tgcgatctgc atctcaatta
       61 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
      121 cgcccattct ccgccccatc gctgactaat tttttttatt tatgcagagg ccgaggccgc
      181 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
      241 caaaaagctt acttggtcag cggccgctaa tacgactcac tatagggaca tttgcttctg
      301 acacaactgt gttcactagc aacctcaaac agacaccatg gatctggtgc taaaaagatg
      361 ccttcttcat ttggctgtga taggtgcttt gctggctgtg ggggctacaa aaggccccag
      421 aaaccaggac tggcttggtg tctcaaggca actcagaacc aaagcctgga acaggcagct
      481 gtatccagag tggacagaag cccagagact tgactgctgg agaggtggtc aagtgtccct
      541 caaggtcagt aatgatgggc ctacactgat tggtgcaaat gcctccttct ctattgcctt
      601 gaacttccct ggaagccaaa aggtattgcc agatgggcag gttatctggg tcaacaatac
      661 catcatcaat gggagccagg tgtggggagg acagccagtg tatccccagg aaactgacga
      721 tgcctgcatc ttccctgatg gtggaccttg cccatctggc tcttggtctc agaagagaag
      781 ctttgtttat gtctggaaga cctggggcca atactggcaa gttctagggg gcccagtgtc
      841 tgggctgagc attgggacag gcagggcaat gctgggcaca cacaccatgg aagtgactgt
      901 ctaccatcgc cggggatccc ggagctatgt gcctcttgct cattccagct cagccttcac
      961 cattactgac caggtgcctt tctccgtgag cgtgtcccag ttgcgggcct tggatggagg
     1021 gaacaagcac ttcctgagaa atcagcctct gacctttgcc ctccagctcc atgaccccag
     1081 tggctatctg gctgaagctg acctctccta cacctgggac tttggagaca gtagtggaac
     1141 cctgatctct cgggcacttg tggtcactca tacttacctg gagcctggcc cagtcactgc
     1201 ccaggtggtc ctgcaggctg ccattcctct cacctcctgt ggctcctccc cagttccagg
     1261 caccacagat gggcacaggc caactgcaga ggcccctaac accacagctg gccaagtgcc
     1321 tactacagaa gttgtgggta ctacacctgg tcaggcgcca actgcagagc cctctggaac
     1381 cacatctgtg caggtgccaa ccactgaagt cataagcact gcacctgtgc agatgccaac
     1441 tgcagagagc acaggtatga cacctgagaa ggtgccagtt tcagaggtca tgggtaccac
     1501 actggcagag atgtcaactc cagaggctac aggtatgaca cctgcagagg tatcaattgt
     1561 ggtgctttct ggaaccacag ctgcacaggt aacaactaca gagtgggtgg agaccacagc
     1621 tagagagcta cctatccctg agcctgaagg tccagatgcc agctcaatca tgtctacgga
     1681 aagtattaca ggttccctgg gccccctgct ggatggtaca gccaccttaa ggctggtgaa
     1741 gagacaagtc cccctggatt gtgttctgta tcgatatggt tccttttccg tcaccctgga
     1801 cattgtccag ggtattgaaa gtgccgagat cctgcaggct gtgccgtccg gtgaggggga
     1861 tgcatttgag ctgactgtgt cctgccaagg cgggctgccc aaggaagcct gcatggagat
     1921 ctcatcgcca gggtgccagc cccctgccca gcggctgtgc cagcctgtgc tacccagccc
     1981 agcctgccag ctggttctgc accagatact gaagggtggc tcggggacat actgcctcaa
     2041 tgtgtctctg gctgatacca acagcctggc agtggtcagc acccagctta tcatgcctgg
     2101 tcaagaagca ggccttgggc aggttccgct gatcgtgggc atcttgctgg tgttgatggc
     2161 tgtggtcctt gcatctctga tatataggcg cagacttatg aagcaagact tctccgtacc
     2221 ccagttgcca catagcagca gtcactggct gcgtctaccc cgcatcttct gctcttgtcc
     2281 cattggtgag aacagccccc tcctcagtgg gcagcaggtc tgagctcgct ttcttgctgt
     2341 ccaatttcta ttaaaggttc ctttgttccc taagtccaac tactaaactg ggggatatta
     2401 tgaagggcct tgagcatctg gattctgcct aataaaaaac atttattttc attgcaagaa
     2461 ttcccgggcc cgcggatcca tgagctagag tcggggcggc cggccgcttc gagcagacat
     2521 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     2581 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     2641 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     2701 tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc gataaggatc tgaacgatgg
     2761 agcggagaat gggcggaact gggcggagtt aggggcggga tgggcggagt taggggcggg
     2821 actatggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct
     2881 ggggactttc cacacctggt tgctgactaa ttgagatgca tgctttgcat acttctgcct
     2941 gctggggagc ctggggactt tccacaccct aactgacaca cattccacag cggatccgtc
     3001 gaccgatgcc cttgagagcc ttcaacccag tcagctcctt ccggtgggcg cggggcatga
     3061 ctatcgtcgc cgcacttatg actgtcttct ttatcatgca actcgtagga caggtgccgg
     3121 cagcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     3181 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     3241 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     3301 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     3361 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     3421 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     3481 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     3541 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     3601 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     3661 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     3721 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     3781 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     3841 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     3901 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     3961 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     4021 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     4081 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     4141 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     4201 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     4261 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     4321 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     4381 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     4441 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     4501 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     4561 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     4621 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     4681 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     4741 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     4801 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     4861 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     4921 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     4981 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     5041 ccccgaaaag tgccacctga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     5101 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     5161 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     5221 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     5281 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     5341 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     5401 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     5461 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttgccat
     5521 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     5581 cgccagccca agctaccatg ataagtaagt aatattaagg tacgggaggt acttggagcg
     5641 gccgcaataa aatatcttta ttttcattac atctgtgtgt tggttttttg tgtgaatcga
     5701 tagtactaac atacgctctc catcaaaaca aaacgaaaca aaacaaacta gcaaaatagg
     5761 ctgtccccag tgcaagtgca ggtgccagaa catttctcta tcgata
//
