LOCUS       sequence-173039-        6330 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Membrane Receptor, C1 Cloning Vector, Excitation:N/A, 
            Emission:N/A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6330)
  TITLE     Michael Davidson Unpublished
REFERENCE   2  (bases 1 to 6330)
  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: TBRII-C-18
FEATURES             Location/Qualifiers
     source          1..6330
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      243..252
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             249..794
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="fast-reacting variant, optimized for mammalian 
                     expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     CDS             2176..2187
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     polyA_signal    2754..2875
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2791..2810)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2845..2864
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2882..3337)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3019..3040)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3231..3250
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3364..3468
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3470..3827
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3470..3661
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3493..3513)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3678..3813
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3862..4656
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3862..4656
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3916..3935)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4526..4545
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4844..4863)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4888..4935
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5264..5852
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5753..5772
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        6027..6330
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg
      241 tcgccaccat ggacaaagac tgcgaaatga agcgcaccac cctggatagc cctctgggca
      301 agctggaact gtctgggtgc gaacagggcc tgcaccgtat catcttcctg ggcaaaggaa
      361 catctgccgc cgacgccgtg gaagtgcctg ccccagccgc cgtgctgggc ggaccagagc
      421 cactgatgca ggccaccgcc tggctcaacg cctactttca ccagcctgag gccatcgagg
      481 agttccctgt gccagccctg caccacccag tgttccagca ggagagcttt acccgccagg
      541 tgctgtggaa actgctgaaa gtggtgaagt tcggagaggt catcagctac agccacctgg
      601 ccgccctggc cggcaatccc gccgccaccg ccgccgtgaa aaccgccctg agcggaaatc
      661 ccgtgcccat tctgatcccc tgccaccggg tggtgcaggg cgacctggac gtggggggct
      721 acgagggcgg gctcgccgtg aaagagtggc tgctggccca cgagggccac agactgggca
      781 agcctgggct gggttccgga ctcagatctc gagagagtgg ctctggtggg tcgagtggaa
      841 gtggcagcgg tcgggggctg ctcaggggcc tgtggccgct gcacatcgtc ctgtggacgc
      901 gtatcgccag cacgatccca ccgcacgttc agaagtcgga tgtggaaatg gaggcccaga
      961 aagatgaaat catctgcccc agctgtaata ggactgccca tccactgaga catattaata
     1021 acgacatgat agtcactgac aacaacggtg cagtcaagtt tccacaactg tgtaaatttt
     1081 gtgatgtgag attttccacc tgtgacaacc agaaatcctg catgagcaac tgcagcatca
     1141 cctccatctg tgagaagcca caggaagtct gtgtggctgt atggagaaag aatgacgaga
     1201 acataacact agagacagtt tgccatgacc ccaagctccc ctaccatgac tttattctgg
     1261 aagatgctgc ttctccaaag tgcattatga aggaaaaaaa aaagcctggt gagactttct
     1321 tcatgtgttc ctgtagctct gatgagtgca atgacaacat catcttctca gaagaatata
     1381 acaccagcaa tcctgacttg ttgctagtca tatttcaagt gacaggcatc agcctcctgc
     1441 caccactggg agttgccata tctgtcatca tcatcttcta ctgctaccgc gttaaccggc
     1501 agcagaagct gagttcaacc tgggaaaccg gcaagacgcg gaagctcatg gagttcagcg
     1561 agcactgtgc catcatcctg gaagatgacc gctctgacat cagctccacg tgtgccaaca
     1621 acatcaacca caacacagag ctgctgccca ttgagctgga caccctggtg gggaaaggtc
     1681 gctttgctga ggtctataag gccaagctga agcagaacac ttcagagcag tttgagacag
     1741 tggcagtcaa gatctttccc tatgaggagt atgcctcttg gaagacagag aaggacatct
     1801 tctcagacat caatctgaag catgagaaca tactccagtt cctgacggct gaggagcgga
     1861 agacggagtt ggggaaacaa tactggctga tcaccgcctt ccacgccaag ggcaacctac
     1921 aggagtacct gacgcggcat gtcatcagct gggaggacct gcgcaagctg ggcagctccc
     1981 tcgcccgggg gattgctaac ctccacagtg atcacactcc atgtgggagg cccaagatgc
     2041 ccatcgtgca cagggacctc aagagctcca atatcctcgt gaagaacgac ctaacctgct
     2101 gcctgtgtga ctttgggctt tccctgcgtc tggaccctac tctgtctgtg gatgacctgg
     2161 ctaacagtgg gcaggtggga actgcaagat acatggctcc agaagtccta gaatccagga
     2221 tgaatttgga gaatgttgag tccttcaagc agaccgatgt ctactccatg gctctggtgc
     2281 tctgggaaat gacatctcgc tgtaatgcag tgggagaagt aaaagattat gagcctccat
     2341 ttggttccaa ggtgcgggag cacccctgtg tcgaaagcat gaaggacaac gtgttgagag
     2401 atcgagggcg accagaaatt cccagcttct ggctcaacca ccagggcatc cagatggtgt
     2461 gtgagacgtt gactgagtgc tgggaccacg acccagaggc ccgtctcaca gcccagtgtg
     2521 tggcagaacg cttcagtgag ctggagcatc tggacaggct ctcggggagg agctgctcgg
     2581 aggagaagat tcctgaagac ggctccctaa acactaccaa atagggatcc accggatcta
     2641 gataactgat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     2701 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     2761 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     2821 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttaacgcg
     2881 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     2941 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     3001 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     3061 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     3121 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     3181 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     3241 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     3301 acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat
     3361 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     3421 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa
     3481 agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     3541 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     3601 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     3661 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     3721 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     3781 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag
     3841 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     3901 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     3961 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     4021 tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg
     4081 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     4141 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     4201 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     4261 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     4321 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     4381 ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     4441 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     4501 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     4561 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     4621 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     4681 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     4741 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     4801 atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga
     4861 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg
     4921 ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc
     4981 caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc
     5041 caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag
     5101 cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     5161 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     5221 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     5281 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     5341 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     5401 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     5461 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     5521 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     5581 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     5641 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     5701 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     5761 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     5821 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     5881 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     5941 tggataaccg tattaccgcc atgcattagt tattaatagt aatcaattac ggggtcatta
     6001 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
     6061 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
     6121 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
     6181 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
     6241 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     6301 atctacgtat tagtcatcgc tattaccatg
//
