LOCUS       sequence-433267-        5329 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector to transiently express full-length human
            Tspan12 with a Rho1D4 tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5329)
  AUTHORS   Bruguera ES, Mahoney JP, Weis WI
  TITLE     The co-receptor Tspan12 directly captures Norrin to promote 
            ligand-specific beta-catenin signaling
  JOURNAL   bioRxiv 2024.02.03.578714
REFERENCE   2  (bases 1 to 5329)
  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: "bioRxiv 
            2024.02.03.578714"
COMMENT     Sequence Label: hTspan12(1-305)-1D4_pTT5
FEATURES             Location/Qualifiers
     source          1..5329
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     14..31
                     /label=L4440
                     /note="L4440 vector, forward primer"
     enhancer        78..457
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        458..661
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     612..632
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     658..682
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     1163..1182
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1226..1235
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2241..2296
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2295..2314)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3379..3397)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3465..3569
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3570..3638,3639..4430)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3788..3807)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4601..5189
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5090..5109
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacgaccgag cgcagcgagt cagtgagcga ggaagcgtac atttatattg gctcatgtcc
       61 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      121 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      181 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      241 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      301 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      361 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      421 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      481 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      541 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      601 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      661 tcgtttagtg aaccgtcaga tcctcactct cttccgcatc gctgtctgcg agggccagct
      721 gttgggctcg cggttgagga caaactcttc gcggtctttc cagtactctt ggatcggaaa
      781 cccgtcggcc tccgaacggt actccgccac cgagggacct gagcgagtcc gcatcgaccg
      841 gatcggaaaa cctctcgaga aaggcgtcta accagtcaca gtcgcaaggt aggctgagca
      901 ccgtggcggg cggcagcggg tggcggtcgg ggttgtttct ggcggaggtg ctgctgatga
      961 tgtaattaaa gtaggcggtc ttgagacggc ggatggtcga ggtgaggtgt ggcaggcttg
     1021 agatccagct gttggggtga gtactccctc tcaaaagcgg gcattacttc tgcgctaaga
     1081 ttgtcagttt ccaaaaacga ggaggatttg atattcacct ggcccgatct ggccatacac
     1141 ttgagtgaca atgacatcca ctttgccttt ctctccacag gtgtccactc ccaggtccaa
     1201 gtttaaacgg atctctagcg aattcgccac catggccaga gaagattccg tgaagtgtct
     1261 gcgctgcctg ctctacgccc tcaatctgct cttttggtta atgtccatca gtgtgttggc
     1321 agtttctgct tggatgaggg actacctaaa taatgttctc actttaactg cagaaacgag
     1381 ggtagaggaa gcagtcattt cgacttactt tcctgtggtt catccggtca tgattgctgt
     1441 ttgctgtttc cttatcattg tggggatgtt aggatattgt ggaacggtga aaagaaatct
     1501 gttgcttctt gcatggtact ttggaagttt gcttgtcatt ttctgtgtag aactggcttg
     1561 tggcgtttgg acatatgaac aggaacttat ggttccagta caatggtcag atatggtcac
     1621 tttgaaagcc aggatgacaa attatggatt acctagatat cggtggctta ctcatgcttg
     1681 gaattttttt cagagagagt ttaagtgctg tggagtagta tatttcactg actggttgga
     1741 aatgacagag atggactggc ccccagattc ctgctgtgtt agagaattcc caggatgttc
     1801 caaacaggcc caccaggaag atctcagtga cctttatcaa gagggttgtg ggaagaaaat
     1861 gtattccttt ttgagaggaa ccaaacaact gcaggtgctg aggtttctgg gaatctccat
     1921 tggggtgaca caaatcctgg ccatgattct caccattact ctgctctggg ctctgtatta
     1981 tgatagaagg gagcctggga cagaccaaat gatgtccttg aagaatgaca actctcagca
     2041 cctgtcatgt ccctcagtag aactgttgaa accaagcctg tcaagaatct ttgaacacac
     2101 atccatggca aacagcttta atacacactt tgagatggag gagttaggta ccgagaccag
     2161 ccaggtggcc cctgcataaa agcttgctag cggccgctcg aggccggcaa ggccggatcc
     2221 cccgacctcg acctctggct aataaaggaa atttattttc attgcaatag tgtgttggaa
     2281 ttttttgtgt ctctcactcg gaaggacata tgggagggca aatcatttgg tcgagatccc
     2341 tcggagatct ctagctagag gatcgatccc cgccccggac gaactaaacc tgactacgac
     2401 atctctgccc cttcttcgcg gggcagtgca tgtaatccct tcagttggtt ggtacaactt
     2461 gccaactgaa ccctaaacgg gtagcatatg cttcccgggt agtagtatat actatccaga
     2521 ctaaccctaa ttcaatagca tatgttaccc aacgggaagc atatgctatc gaattagggt
     2581 tagtaaaagg gtcctaagga acagcgatgt aggtgggcgg gccaagatag gggcgcgatt
     2641 gctgcgatct ggaggacaaa ttacacacac ttgcgcctga gcgccaagca cagggttgtt
     2701 ggtcctcata ttcacgaggt cgctgagagc acggtgggct aatgttgcca tgggtagcat
     2761 atactaccca aatatctgga tagcatatgc tatcctaatc tatatctggg tagcataggc
     2821 tatcctaatc tatatctggg tagcatatgc tatcctaatc tatatctggg tagtatatgc
     2881 tatcctaatt tatatctggg tagcataggc tatcctaatc tatatctggg tagcatatgc
     2941 tatcctaatc tatatctggg tagtatatgc tatcctaatc tgtatccggg tagcatatgc
     3001 tatcctaata gagattaggg tagtatatgc tatcctaatt tatatctggg tagcatatac
     3061 tacccaaata tctggatagc atatgctatc ctaatctata tctgggtagc atatgctatc
     3121 ctaatctata tctgggtagc ataggctatc ctaatctata tctgggtagc atatgctatc
     3181 ctaatctata tctgggtagt atatgctatc ctaatttata tctgggtagc ataggctatc
     3241 ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagt atatgctatc
     3301 ctaatctgta tccgggtagc atatgctatc ctcatgataa gctgtcaaac atgagaatta
     3361 attcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     3421 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     3481 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     3541 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     3601 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     3661 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     3721 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     3781 aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc
     3841 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     3901 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     3961 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     4021 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     4081 taccaaacga cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac
     4141 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     4201 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     4261 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     4321 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     4381 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     4441 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     4501 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     4561 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     4621 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     4681 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     4741 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     4801 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     4861 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     4921 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     4981 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     5041 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     5101 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     5161 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     5221 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     5281 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccg
//
