LOCUS       sequence-471882-        6116 bp    DNA     circular SYN 14-JUL-2025
DEFINITION  Express TGFBR1-NanoLuc(R) Fusion Protein in Mammalian Cells under a 
            CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6116)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 6116)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: TGFBR1-NanoLuc Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6116
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          857..989
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     972..991
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1033..1052
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1033..1051
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2595..3107
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(3195..3213)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3250..3371
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3261..3280)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3315..3334
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3616..3973
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3616..3807
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3639..3659)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3824..3959
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3988..4035
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4067..4861
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4121..4140)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4731..4750
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4925..4973
                     /note="synthetic polyadenylation signal"
     rep_origin      5130..5675
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     5576..5595
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5792..5809
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      721 tggtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
      841 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
      901 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
      961 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1021 aggctagagt attaatacga ctcactatag ggctagcgat cgccatggag gcggcggtcg
     1081 ctgctccgcg tccccggctg ctcctcctcg tgctggcggc ggcggcggcg gcggcggcgg
     1141 cgctgctccc gggggcgacg gcgttacagt gtttctgcca cctctgtaca aaagacaatt
     1201 ttacttgtgt gacagatggg ctctgctttg tctctgtcac agagaccaca gacaaagtta
     1261 tacacaacag catgtgtata gctgaaattg acttaattcc tcgagatagg ccgtttgtat
     1321 gtgcaccctc ttcaaaaact gggtctgtga ctacaacata ttgctgcaat caggaccatt
     1381 gcaataaaat agaacttcca actactgtaa agtcatcacc tggccttggt cctgtggaac
     1441 tggcagctgt cattgctgga ccagtgtgct tcgtctgcat ctcactcatg ttgatggtct
     1501 atatctgcca caaccgcact gtcattcacc atcgagtgcc aaatgaagag gacccttcat
     1561 tagatcgccc ttttatttca gagggtacta cgttgaaaga cttaatttat gatatgacaa
     1621 cgtcaggttc tggctcaggt ttaccattgc ttgttcagag aacaattgcg agaactattg
     1681 tgttacaaga aagcattggc aaaggtcgat ttggagaagt ttggagagga aagtggcggg
     1741 gagaagaagt tgctgttaag atattctcct ctagagaaga acgttcgtgg ttccgtgagg
     1801 cagagattta tcaaactgta atgttacgtc atgaaaacat cctgggattt atagcagcag
     1861 acaataaaga caatggtact tggactcagc tctggttggt gtcagattat catgagcatg
     1921 gatccctttt tgattactta aacagataca cagttactgt ggaaggaatg ataaaacttg
     1981 ctctgtccac ggcgagcggt cttgcccatc ttcacatgga gattgttggt acccaaggaa
     2041 agccagccat tgctcataga gatttgaaat caaagaatat cttggtaaag aagaatggaa
     2101 cttgctgtat tgcagactta ggactggcag taagacatga ttcagccaca gataccattg
     2161 atattgctcc aaaccacaga gtgggaacaa aaaggtacat ggcccctgaa gttctcgatg
     2221 attccataaa tatgaaacat tttgaatcct tcaaacgtgc tgacatctat gcaatgggct
     2281 tagtattctg ggaaattgct cgacgatgtt ccattggtgg aattcatgaa gattaccaac
     2341 tgccttatta tgatcttgta ccttctgacc catcagttga agaaatgaga aaagttgttt
     2401 gtgaacagaa gttaaggcca aatatcccaa acagatggca gagctgtgaa gccttgagag
     2461 taatggctaa aattatgaga gaatgttggt atgccaatgg agcagctagg cttacagcat
     2521 tgcggattaa gaaaacatta tcgcaactca gtcaacagga aggcatcaaa atggtttctc
     2581 tcggctcgag cggcgtcttc acactcgaag atttcgttgg ggactggcga cagacagccg
     2641 gctacaacct ggaccaagtc cttgaacagg gaggtgtgtc cagtttgttt cagaatctcg
     2701 gggtgtccgt aactccgatc caaaggattg tcctgagcgg tgaaaatggg ctgaagatcg
     2761 acatccatgt catcatcccg tatgaaggtc tgagcggcga ccaaatgggc cagatcgaaa
     2821 aaatttttaa ggtggtgtac cctgtggatg atcatcactt taaggtgatc ctgcactatg
     2881 gcacactggt aatcgacggg gttacgccga acatgatcga ctatttcgga cggccgtatg
     2941 aaggcatcgc cgtgttcgac ggcaaaaaga tcactgtaac agggaccctg tggaacggca
     3001 acaaaattat cgacgagcgc ctgatcaacc ccgacggctc cctgctgttc cgagtaacca
     3061 tcaacggagt gaccggctgg cggctgtgcg aacgcattct ggcgtaaggc cgcgactcta
     3121 gagtcgacct gcaggcatgc aagctgatcc ggctgctaac aaagcccgaa aggaagctga
     3181 gttggctgct gccaccgctg agcaataact agcataaccc cttggggcgg ccgcttcgag
     3241 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     3301 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     3361 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggagatgt
     3421 gggaggtttt tttaagcaag taaaacctct acaaatgtgg taaaatcgaa ttttaacaaa
     3481 atattaacgc ttacaatttc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     3541 cacaccgcat acgcggatct gcgcagcacc atggcctgaa ataacctctg aaagaggaac
     3601 ttggttaggt accttctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg
     3661 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3721 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3781 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     3841 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     3901 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     3961 aggcttttgc aaaaagctta attaactgtt gacaattaat catcggcata gtatatcggc
     4021 atagtataat acgacaaggt gaggaactaa acccaggagg cagatcatga ttgaacaaga
     4081 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     4141 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     4201 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     4261 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     4321 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     4381 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     4441 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     4501 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     4561 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     4621 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     4681 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     4741 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     4801 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     4861 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgatgg
     4921 ccgcaataaa atatctttat tttcattaca tctgtgtgtt ggttttttgt gtgaatcgat
     4981 agcgataagg atccgctcat gcgatgataa gctgtcaaac atgagaatta caacttatat
     5041 cgtatggggc tgacttcagg tgctacattt gaagagataa attgcactga aatctagaaa
     5101 tattttatct gattaataag atgatcttct tgagatcgtt ttggtctgcg cgtaatctct
     5161 tgctctgaaa acgaaaaaac cgccttgcag ggcggttttt cgaaggttct ctgagctacc
     5221 aactctttga accgaggtaa ctggcttgga ggagcgcagt caccaaaact tgtcctttca
     5281 gtttagcctt aaccggcgca tgacttcaag actaactcct ctaaatcaat taccagtggc
     5341 tgctgccagt ggtgcttttg catgtctttc cgggttggac tcaagacgat agttaccgga
     5401 taaggcgcag cggtcggact gaacgggggg ttcgtgcata cagtccagct tggagcgaac
     5461 tgcctacccg gaactgagtg tcaggcgtgg aatgagacaa acgcggccat aacagcggaa
     5521 tgacaccggt aaaccgaaag gcaggaacag gagagcgcac gagggagccg ccagggggaa
     5581 acgcctggta tctttatagt cctgtcgggt ttcgccacca ctgatttgag cgtcagattt
     5641 cgtgatgctt gtcagggggg cggagcctat ggaaaaacgg ctttgccgcg gccctctcac
     5701 ttccctgtta agtatcttcc tggcatcttc caggaaatct ccgccccgtt cgtaagccat
     5761 ttccgctcgc cgcagtcgaa cgaccgagcg tagcgagtca gtgagcgagg aagcggaata
     5821 tatcctgtat cacatattct gctgacgcac cggtgcagcc ttttttctcc tgccacatga
     5881 agcacttcac tgacaccctc atcagtgcca acatagtaag ccagtataca ctccgctagc
     5941 gctgatgtcc ggcggtgctt ttgccgttac gcaccacccc gtcatgagat tacgcgtttt
     6001 agcggtgcgt acaattaagg gattatggta aatccactta ctgtctgccc tcgtagccat
     6061 cgagataaac cgcagtactc cggccacgat gcgtccggcg tagaggatcg agatct
//
