LOCUS       sequence-477826-        5821 bp    DNA     circular SYN 07-JUL-2025
DEFINITION  Template for in vitro transcription of PMEL (gp100), with signal 
            peptide replaced by the IL6 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 5821)
  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 5821)
  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: pIL6-PMEL-hbb
FEATURES             Location/Qualifiers
     source          1..5821
                     /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    2538..2659
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2549..2568)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2603..2622
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2981..3001
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     complement(3153..3170)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3324..3912)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3404..3423)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4083..4874,4875..4943))
                     /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     4706..4725
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4944..5048)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5075..5530
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5162..5181)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5372..5393
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5661..5709
                     /note="synthetic polyadenylation signal"
     misc_feature    5723..5814
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5763..5782
                     /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 gactccttct ccacaagcgc
      361 cttcggtcca gttgccttct ccctgggcct gctcctggtg ttgcctgctg ccttccctgc
      421 cccaaaaggc cccagaaacc aggactggct tggtgtctca aggcaactca gaaccaaagc
      481 ctggaacagg cagctgtatc cagagtggac agaagcccag agacttgact gctggagagg
      541 tggtcaagtg tccctcaagg tcagtaatga tgggcctaca ctgattggtg caaatgcctc
      601 cttctctatt gccttgaact tccctggaag ccaaaaggta ttgccagatg ggcaggttat
      661 ctgggtcaac aataccatca tcaatgggag ccaggtgtgg ggaggacagc cagtgtatcc
      721 ccaggaaact gacgatgcct gcatcttccc tgatggtgga ccttgcccat ctggctcttg
      781 gtctcagaag agaagctttg tttatgtctg gaagacctgg ggccaatact ggcaagttct
      841 agggggccca gtgtctgggc tgagcattgg gacaggcagg gcaatgctgg gcacacacac
      901 catggaagtg actgtctacc atcgccgggg atcccggagc tatgtgcctc ttgctcattc
      961 cagctcagcc ttcaccatta ctgaccaggt gcctttctcc gtgagcgtgt cccagttgcg
     1021 ggccttggat ggagggaaca agcacttcct gagaaatcag cctctgacct ttgccctcca
     1081 gctccatgac cccagtggct atctggctga agctgacctc tcctacacct gggactttgg
     1141 agacagtagt ggaaccctga tctctcgggc acttgtggtc actcatactt acctggagcc
     1201 tggcccagtc actgcccagg tggtcctgca ggctgccatt cctctcacct cctgtggctc
     1261 ctccccagtt ccaggcacca cagatgggca caggccaact gcagaggccc ctaacaccac
     1321 agctggccaa gtgcctacta cagaagttgt gggtactaca cctggtcagg cgccaactgc
     1381 agagccctct ggaaccacat ctgtgcaggt gccaaccact gaagtcataa gcactgcacc
     1441 tgtgcagatg ccaactgcag agagcacagg tatgacacct gagaaggtgc cagtttcaga
     1501 ggtcatgggt accacactgg cagagatgtc aactccagag gctacaggta tgacacctgc
     1561 agaggtatca attgtggtgc tttctggaac cacagctgca caggtaacaa ctacagagtg
     1621 ggtggagacc acagctagag agctacctat ccctgagcct gaaggtccag atgccagctc
     1681 aatcatgtct acggaaagta ttacaggttc cctgggcccc ctgctggatg gtacagccac
     1741 cttaaggctg gtgaagagac aagtccccct ggattgtgtt ctgtatcgat atggttcctt
     1801 ttccgtcacc ctggacattg tccagggtat tgaaagtgcc gagatcctgc aggctgtgcc
     1861 gtccggtgag ggggatgcat ttgagctgac tgtgtcctgc caaggcgggc tgcccaagga
     1921 agcctgcatg gagatctcat cgccagggtg ccagccccct gcccagcggc tgtgccagcc
     1981 tgtgctaccc agcccagcct gccagctggt tctgcaccag atactgaagg gtggctcggg
     2041 gacatactgc ctcaatgtgt ctctggctga taccaacagc ctggcagtgg tcagcaccca
     2101 gcttatcatg cctggtcaag aagcaggcct tgggcaggtt ccgctgatcg tgggcatctt
     2161 gctggtgttg atggctgtgg tccttgcatc tctgatatat aggcgcagac ttatgaagca
     2221 agacttctcc gtaccccagt tgccacatag cagcagtcac tggctgcgtc taccccgcat
     2281 cttctgctct tgtcccattg gtgagaacag ccccctcctc agtgggcagc aggtctgagc
     2341 tcgctttctt gctgtccaat ttctattaaa ggttcctttg ttccctaagt ccaactacta
     2401 aactggggga tattatgaag ggccttgagc atctggattc tgcctaataa aaaacattta
     2461 ttttcattgc aagaattccc gggcccgcgg atccatgagc tagagtcggg gcggccggcc
     2521 gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     2581 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     2641 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     2701 ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta aaatcgataa
     2761 ggatctgaac gatggagcgg agaatgggcg gaactgggcg gagttagggg cgggatgggc
     2821 ggagttaggg gcgggactat ggttgctgac taattgagat gcatgctttg catacttctg
     2881 cctgctgggg agcctgggga ctttccacac ctggttgctg actaattgag atgcatgctt
     2941 tgcatacttc tgcctgctgg ggagcctggg gactttccac accctaactg acacacattc
     3001 cacagcggat ccgtcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt
     3061 gggcgcgggg catgactatc gtcgccgcac ttatgactgt cttctttatc atgcaactcg
     3121 taggacaggt gccggcagcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     3181 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     3241 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     3301 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     3361 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     3421 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     3481 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     3541 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     3601 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     3661 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgtt
     3721 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     3781 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     3841 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     3901 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     3961 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     4021 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     4081 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     4141 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     4201 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     4261 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     4321 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     4381 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     4441 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     4501 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     4561 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     4621 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     4681 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     4741 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     4801 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     4861 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     4921 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     4981 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     5041 gttccgcgca catttccccg aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc
     5101 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     5161 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     5221 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     5281 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     5341 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     5401 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     5461 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     5521 cttacaattt gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     5581 gcctcttcgc tattacgcca gcccaagcta ccatgataag taagtaatat taaggtacgg
     5641 gaggtacttg gagcggccgc aataaaatat ctttattttc attacatctg tgtgttggtt
     5701 ttttgtgtga atcgatagta ctaacatacg ctctccatca aaacaaaacg aaacaaaaca
     5761 aactagcaaa ataggctgtc cccagtgcaa gtgcaggtgc cagaacattt ctctatcgat
     5821 a
//
