LOCUS       sequence-412364-        6124 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GPNMB protein expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6124)
  TITLE     TREAT-AD plasmids Jun 2023
REFERENCE   2  (bases 1 to 6124)
  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"
COMMENT     Sequence Label: GPNMBA-c200
FEATURES             Location/Qualifiers
     source          1..6124
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          640..1656
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1580..1602
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1664..1683
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1738..1747
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3289..3333
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             3346..3363
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3475..3494)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3540..3595
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3594..3613)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3956..3972)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3956..3972)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3969..3991)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3980..3996
                     /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(4004..4010,4011..4028,4029..4034))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4049..4070
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4199..4216)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4370..4958)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4450..4469)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5129..5920,5921..5989))
                     /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     5752..5771
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5990..6094)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
      421 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
      481 tgggggcggg gggggggggg gggggcgcgc gggggggggg gsggggcgrg gcggagaggt
      541 gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg
      601 cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc gcgctgcctt
      661 cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc cccggctctg actgaccgcg
      721 ttactcccac aggtgagcgg gcgggacggc ccttctcctc cgggctgtaa ttagcgcttg
      781 gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa gccttgaggg gctccgggag
      841 ggccctttgt gcggggggag cggctcgggg ggtgcgtgcg tgtgtgtgtg cgtggggagc
      901 gccgcgtgcg gctccgcgct gcccggcggc tgtgagcgct gcgggcgcgg cgcggggctt
      961 tgtgcgctcc gcagtgtgcg cgaggggagc gcggccgggg gcggtgcccc gcggtgcggg
     1021 gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt gcgtgggggg gtgagcaggg
     1081 ggtgtgggcg cgtcggtcgg gctgcaaccc cccctgcacc cccctccccg agttgctgag
     1141 cacggcccgg cttcgggtgc ggggctccgt acggggcgtg gcgcggggct cgccgtgccg
     1201 ggcggggggt ggcggcaggt gggggtgccg ggcggggcgg ggccgcctcg ggccggggag
     1261 ggctcggggg aggggcgcgg cggcccccgg agcgccggcg gctgtcgagg cgcggcgagc
     1321 cgcagccatt gccttttatg gtaatcgtgc gagagggcgc agggacttcc tttgtcccaa
     1381 atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc cctctagcgg gcgcggggcg
     1441 aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg gccttcgtgc gtcgccgcgc
     1501 cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc ggggggacgg ctgccttcgg
     1561 gggggacggg gcagggcggg gttcggcttc tggcgtgtga ccggcggctc tagagcctct
     1621 gctaaccatg ttcatgcctt cttctttttc ctacagctcc tgggcaacgt gctggttatt
     1681 gtgctgtctc atcattttgg caaagaattg cggccgtctc aggccgaatt caagcttgcc
     1741 accatgggga tccttcccag ccctgggatg cctgcgctgc tctccctcgt gagccttctc
     1801 tccgtgctgc tgatgggttg cgtagctgaa accggtaaac ggtttcatga tgtactcggt
     1861 aacgaaagac cgagcgcata catgagagaa cataatcaac tgaatggatg gtcctcagat
     1921 gagaacgatt ggaatgaaaa gttgtaccca gtatggaaaa ggggtgatat gagatggaag
     1981 aattcatgga aaggcggcag agtccaagcg gttctgacat cagacagtcc cgcgctggtt
     2041 ggcagtaaca taacttttgc ggtgaatctg atcttccccc gctgccaaaa ggaagatgct
     2101 aacgggaaca tcgtttatga gaagaattgt aggaatgaag ctggcctttc tgccgatccc
     2161 tacgtgtata attggacggc gtggagcgaa gattcagatg gcgaaaatgg gacggggcag
     2221 tcacatcata atgtctttcc cgatggcaag ccttttccgc atcaccccgg ctggcgtcgc
     2281 tggaacttta tttacgtctt tcataccctc ggacaatatt ttcagaagct ggggcgttgt
     2341 agcgtccgag tttccgtcaa tactgcaaat gtcactcttg gcccccagct catggaagtg
     2401 actgtctacc ggcgtcatgg ccgggcgtat gtgcccatag ctcaagttaa ggatgtgtac
     2461 gtagtcaccg accagatccc cgttttcgta actatgtttc agaagaacga cagaaactct
     2521 agcgatgaaa cattcctgaa ggatcttcca ataatgtttg acgttcttat tcacgatccg
     2581 tcccacttct tgaattatag taccatcaat tataagtggt catttggtga caataccggg
     2641 ctgtttgtgt ctactaatca tacagtcaat catacttacg tcctgaacgg gacgttctcc
     2701 ctgaacctta cggtaaaagc tgctgctcca ggcccctgtc caccgccccc acctcctccg
     2761 aggccttcca agcccacgcc atctctggct accactctta agtcatatga ctccaatacc
     2821 cccggaccag caggtgataa tcctctggag ttgagcagga ttcccgatga aaattgccag
     2881 atcaaccgat acgggcattt tcaagccaca atcacgatag tagaaggaat cctggaagtc
     2941 aacattattc aaatgacaga cgtcctgatg cccgtaccat ggcccgaatc ttcactgata
     3001 gactttgtag tcacatgcca aggatcaatt ccgactgaag tctgcacaat catctcagat
     3061 ccaacctgtg agataactca aaatacagtg tgctcccctg tggatgtaga tgaaatgtgc
     3121 ttgcttactg tcaggagaac ttttaacggc tccggcacgt actgtgtcaa tctcactctc
     3181 ggggatgaca ctagccttgc tctgacttcc actctcatct cagtacccga ccgagaccca
     3241 gcctcacctc tccggatggc taacggtacc ggaggttccg gtggttccgg tctgaatgat
     3301 atctttgaag ctcagaagat tgaatggcat gaaggacgta ccaagcacca ccatcaccat
     3361 cactaatgat cactcgagac tagtatcgcg ataattcact cctcaggtgc aggctgccta
     3421 tcagaaggtg gtggctggtg tggccaatgc cctggctcac aaataccact gagatctttt
     3481 tccctctgcc aaaaattatg gggacatcat gaagcccctt gagcatctga cttctggcta
     3541 ataaaggaaa tttattttca ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg
     3601 aaggacatat gggagggcaa atcatttaaa acatcagaat gagtatttgg tttagagttt
     3661 ggcaacatat gcccatatgc tggctgccat gaacaaaggt tggctataaa gaggtcatca
     3721 gtatatgaaa cagccccctg ctgtccattc cttattccat agaaaagcct tgacttgagg
     3781 ttagattttt tttatatttt gttttgtgtt atttttttct ttaacatccc taaaattttc
     3841 cttacatgtt ttactagcca gatttttcct cctctcctga ctactcccag tcatagctgt
     3901 ccctcttctc ttatggagat ccctcgacct gcagcccaag cttggcgtaa tcatggtcat
     3961 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     4021 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     4081 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gcggatcgat ccgctgcatt
     4141 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     4201 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     4261 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     4321 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4381 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4441 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4501 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4561 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4621 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4681 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4741 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4801 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     4861 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     4921 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     4981 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     5041 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     5101 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     5161 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     5221 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     5281 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     5341 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     5401 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     5461 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     5521 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     5581 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     5641 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     5701 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     5761 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     5821 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     5881 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     5941 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     6001 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     6061 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     6121 ggtc
//
