LOCUS       Exported                6315 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Localization: Membrane Receptor, Excitation: N/A, Emission: N/A.
ACCESSION   .
VERSION     .
KEYWORDS    TBRII-N-18
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6315)
  TITLE     Davidson plasmids
REFERENCE   2  (bases 1 to 6315)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6315
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             1331..1342
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     regulatory      1825..1834
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1831..2376
                     /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"
     polyA_signal    2501..2622
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2538..2557)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2592..2611
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2629..3084)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2766..2787)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2978..2997
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3111..3215
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3217..3574
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(3240..3260)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3425..3560
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3487..3506
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3609..4403
                     /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(3663..3682)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4273..4292
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4591..4610)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4635..4682
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5011..5599
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5500..5519
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5774..6077
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6078..6281
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6232..6252
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      6310..4
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
ORIGIN
        1 atgggtcggg ggctgctcag gggcctgtgg ccgctgcaca tcgtcctgtg gacgcgtatc
       61 gccagcacga tcccaccgca cgttcagaag tcggatgtgg aaatggaggc ccagaaagat
      121 gaaatcatct gccccagctg taataggact gcccatccac tgagacatat taataacgac
      181 atgatagtca ctgacaacaa cggtgcagtc aagtttccac aactgtgtaa attttgtgat
      241 gtgagatttt ccacctgtga caaccagaaa tcctgcatga gcaactgcag catcacctcc
      301 atctgtgaga agccacagga agtctgtgtg gctgtatgga gaaagaatga cgagaacata
      361 acactagaga cagtttgcca tgaccccaag ctcccctacc atgactttat tctggaagat
      421 gctgcttctc caaagtgcat tatgaaggaa aaaaaaaagc ctggtgagac tttcttcatg
      481 tgttcctgta gctctgatga gtgcaatgac aacatcatct tctcagaaga atataacacc
      541 agcaatcctg acttgttgct agtcatattt caagtgacag gcatcagcct cctgccacca
      601 ctgggagttg ccatatctgt catcatcatc ttctactgct accgcgttaa ccggcagcag
      661 aagctgagtt caacctggga aaccggcaag acgcggaagc tcatggagtt cagcgagcac
      721 tgtgccatca tcctggaaga tgaccgctct gacatcagct ccacgtgtgc caacaacatc
      781 aaccacaaca cagagctgct gcccattgag ctggacaccc tggtggggaa aggtcgcttt
      841 gctgaggtct ataaggccaa gctgaagcag aacacttcag agcagtttga gacagtggca
      901 gtcaagatct ttccctatga ggagtatgcc tcttggaaga cagagaagga catcttctca
      961 gacatcaatc tgaagcatga gaacatactc cagttcctga cggctgagga gcggaagacg
     1021 gagttgggga aacaatactg gctgatcacc gccttccacg ccaagggcaa cctacaggag
     1081 tacctgacgc ggcatgtcat cagctgggag gacctgcgca agctgggcag ctccctcgcc
     1141 cgggggattg ctaacctcca cagtgatcac actccatgtg ggaggcccaa gatgcccatc
     1201 gtgcacaggg acctcaagag ctccaatatc ctcgtgaaga acgacctaac ctgctgcctg
     1261 tgtgactttg ggctttccct gcgtctggac cctactctgt ctgtggatga cctggctaac
     1321 agtgggcagg tgggaactgc aagatacatg gctccagaag tcctagaatc caggatgaat
     1381 ttggagaatg ttgagtcctt caagcagacc gatgtctact ccatggctct ggtgctctgg
     1441 gaaatgacat ctcgctgtaa tgcagtggga gaagtaaaag attatgagcc tccatttggt
     1501 tccaaggtgc gggagcaccc ctgtgtcgaa agcatgaagg acaacgtgtt gagagatcga
     1561 gggcgaccag aaattcccag cttctggctc aaccaccagg gcatccagat ggtgtgtgag
     1621 acgttgactg agtgctggga ccacgaccca gaggcccgtc tcacagccca gtgtgtggca
     1681 gaacgcttca gtgagctgga gcatctggac aggctctcgg ggaggagctg ctcggaggag
     1741 aagattcctg aagacggctc cctaaacact accaaaagca gcggtggagc aagcgcagcc
     1801 agtggtagcg cggatccacc ggtcgccacc atggacaaag actgcgaaat gaagcgcacc
     1861 accctggata gccctctggg caagctggaa ctgtctgggt gcgaacaggg cctgcaccgt
     1921 atcatcttcc tgggcaaagg aacatctgcc gccgacgccg tggaagtgcc tgccccagcc
     1981 gccgtgctgg gcggaccaga gccactgatg caggccaccg cctggctcaa cgcctacttt
     2041 caccagcctg aggccatcga ggagttccct gtgccagccc tgcaccaccc agtgttccag
     2101 caggagagct ttacccgcca ggtgctgtgg aaactgctga aagtggtgaa gttcggagag
     2161 gtcatcagct acagccacct ggccgccctg gccggcaatc ccgccgccac cgccgccgtg
     2221 aaaaccgccc tgagcggaaa tcccgtgccc attctgatcc cctgccaccg ggtggtgcag
     2281 ggcgacctgg acgtgggggg ctacgagggc gggctcgccg tgaaagagtg gctgctggcc
     2341 cacgagggcc acagactggg caagcctggg ctgggttaag cggccgcgac tctagatcat
     2401 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     2461 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     2521 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     2581 gcattctagt tgtggtttgt ccaaactcat caatgtatct taaggcgtaa attgtaagcg
     2641 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     2701 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     2761 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     2821 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     2881 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     2941 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     3001 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     3061 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     3121 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     3181 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg
     3241 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3301 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     3361 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     3421 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     3481 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     3541 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat
     3601 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     3661 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     3721 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     3781 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     3841 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     3901 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     3961 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     4021 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     4081 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
     4141 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     4201 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     4261 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     4321 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     4381 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     4441 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     4501 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     4561 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     4621 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt
     4681 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     4741 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     4801 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc
     4861 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     4921 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     4981 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     5041 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     5101 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     5161 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     5221 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     5281 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     5341 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     5401 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     5461 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     5521 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     5581 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     5641 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     5701 taccgccatg cattagttat taatagtaat caattacggg gtcattagtt catagcccat
     5761 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     5821 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     5881 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     5941 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     6001 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     6061 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
     6121 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     6181 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
     6241 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga
     6301 tccgctagcg ccacc
//
