LOCUS       Exported                5380 bp ds-DNA     circular SYN 22-MAY-2024
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5380)
  AUTHORS   Department of Anatomy
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, May 22, 2024 from SnapGene 4.3.11
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5380
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     914..931
                     /label=CMV-F
                     /note="color: black; sequence: CGCAAATGGGCGGTAGGCGTG"
     promoter        1007..1025
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     primer_bind     1060..1122
                     /label=EcoRI-NES-FKBP FW
                     /note="color: black; sequence: 
                     TTTGAATTCCTTCAACTTCCTCCTCTTGAACGTCTTACTCTCatgGGAGTGCAGGTG; 
                     added: 2019-06-03"
     CDS             1060..1092
                     /codon_start=1
                     /label=NES
                     /note="color: #993366"
                     /translation="LQLPPLERLTL"
     primer_bind     1096..1127
                     /label=Primer 5
                     /note="color: black; sequence: 
                     tttGAATTCgccaccatgGGAGTGCAGGTGGAAAC"
     regulatory      1102..1111
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
     CDS             1108..1431
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /note="This feature has 2 segments:
                        1: 1108 .. 1110 / #993366
                        2: 1111 .. 1431 / #993366"
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLE"
     primer_bind     1446..1467
                     /label=SphI-CRY2PHR-iLID-LRP6c short FW
                     /note="color: black; sequence: 
                     TTTGCATGCATGGAGTTTCTGGCAACCACA; added: 2019-05-21"
     CDS             1447..1893
                     /codon_start=1
                     /label=iLID
                     /note="color: #0000ff"
                     /translation="MEFLATTLERIEKNFVITDPRLPDNPIIFASDSFLQLTEYSREEI
                     LGRNCRFLQGPETDRATVRKIRDAIDNQTEVTVQLINYTKSGKKFWNVFHLQPMRDYKG
                     DVQYFIGVQLDGTERLHGAAEREAVCLIKKTAFQIAEAANDENYF"
     misc_feature    1450..1893
                     /label=gene synthesis part 756 bp
                     /note="color: #a6acb3"
     primer_bind     complement(1873..1893)
                     /label=PM LRP6c-1 RV
                     /note="color: black; sequence: 
                     tcggtcataaaagtaattttcgtcgttcgc; added: 2019-04-10"
     primer_bind     complement(1878..1905)
                     /label=add-SphI RV
                     /note="color: black; sequence: 
                     gtcataGCATGCaaagtaattttcgTCG; added: 2019-06-10"
     primer_bind     1888..1913
                     /label=add-SphI FW
                     /note="color: black; sequence: TActttGCATGCtatgaccgagccca; 
                     added: 2019-06-10"
     CDS             1900..2364
                     /codon_start=1
                     /product="Homo sapiens"
                     /label=LRP6c
                     /note="color: #ffff00"
                     /translation="YDRAHVTGASSSSSSSTKGTYFPAILNPPPSPATERSHYTMEFGY
                     SSNSPSTHRSYSYRPYSYRHFAPPTTPCSTDVCDSDYAPSRRMTSVATAKGYTSDLNYD
                     SEPVPPPPTPRSQYLSAEENYESCPPSPYTERSYSHHLYPPPPSPCTDSS"
     primer_bind     1900..1923
                     /label=PM LRP6c-1 FW
                     /note="color: black; sequence: 
                     tacttttatgaccgagcccatgttacagga; added: 2019-04-10"
     primer_bind     complement(2269..2283)
                     /label=C term del2 RV
                     /note="color: black; sequence: tctagactagcagctttcatagtt; 
                     added: 2019-04-29"
     primer_bind     complement(2310..2328)
                     /label=C term del1 RV
                     /note="color: black; sequence: agactagtagaggtgatgagaatag; 
                     added: 2019-04-29"
     primer_bind     complement(2340..2364)
                     /label=NheI-LRP6c RV
                     /note="color: black; sequence: 
                     TTTgCTAGCctaggaggagtctgtacagggagag; added: 2019-06-03"
     primer_bind     complement(2343..2361)
                     /label=SphI-LRP6c tandem RV
                     /note="color: black; sequence: 
                     tttGCATGCggaggagtctgtacaggga; added: 2019-04-10"
     primer_bind     complement(2343..2361)
                     /label=tandem LRP6c RV
                     /note="color: black; sequence: 
                     tttACCGGTggaggagtctgtacaggga"
     primer_bind     complement(2355..2378)
                     /label=NheI-CRY2PHR-iLID-LRP6c short RV
                     /note="color: black; sequence: TTTGCTAGctatctagactaggaggag;
                     added: 2019-05-21"
     primer_bind     2361..2382
                     /label=C term del1 FW
                     /note="color: black; sequence: ctctactagtctagatagCTAGCCTCC;
                     added: 2019-04-29"
     primer_bind     2361..2381
                     /label=C term del2 FW
                     /note="color: black; sequence: tgctagtctagatagCTAGcctc; 
                     added: 2019-04-29"
     promoter        complement(2382..2400)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: LEFT"
     polyA_signal    2405..2539
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     promoter        complement(2652..2670)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
                     /note="color: #ffffff; direction: LEFT"
     primer_bind     complement(2691..2707)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     protein_bind    2715..2731
                     /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)."
                     /note="color: #31849b"
     promoter        complement(2739..2769)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This reverse directional feature has 3 segments:
                        1: 2739 .. 2745 / #ffffff / -10
                        2: 2746 .. 2763 / #ffffff
                        3: 2764 .. 2769 / #ffffff / -35"
     protein_bind    2784..2805
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
                     /note="color: #31849b"
     rep_origin      complement(3092..3680)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(3851..4711)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 3851 .. 4642 / #ccffcc
                        2: 4643 .. 4711 / #ccffcc / signal sequence
                     Cleavage site after base 4642"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4712..4816)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
     rep_origin      complement(4842..5297)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        join(5375..5380,1..972)
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
                     /note="color: #ffffff; direction: RIGHT"
ORIGIN
        1 tcaatatggc gtatatggac tcatgccaat tcaatatggt ggatctggac ctgtgccaat
       61 tcaatatggc gtatatggac tcgtgccaat tcaatatggt ggatctggac cccagccaat
      121 tcaatatggc ggacttggca ccatgccaat tcaatatggc ggacctggca ctgtgccaac
      181 tggggagggg tctacttggc acggtgccaa gtttgaggag gggtcttggc cctgtgccaa
      241 gtccgccata ttgaattggc atggtgccaa taatggcggc catattggct atatgccagg
      301 atcaatatat aggcaatatc caatatggcc ctatgccaat atggctattg gccaggttca
      361 atactatgta ttggccctat gccatatagt attccatata tgggttttcc tattgacgta
      421 gatagcccct cccaatgggc ggtcccatat accatatatg gggcttccta ataccgccca
      481 tagccactcc cccattgacg tcaatggtct ctatatatgg tctttcctat tgacgtcata
      541 tgggcggtcc tattgacgta tatggcgcct cccccattga cgtcaattac ggtaaatggc
      601 ccgcctggct caatgcccat tgacgtcaat aggaccaccc accattgacg tcaatgggat
      661 ggctcattgc ccattcatat ccgttctcac gccccctatt gacgtcaatg acggtaaatg
      721 gcccacttgg cagtacatca atatctatta atagtaactt ggcaagtaca ttactattgg
      781 aagtacgcca gggtacattg gcagtactcc cattgacgtc aatggcggta aatggcccgc
      841 gatggctgcc aagtacatcc ccattgacgt caatggggag gggcaatgac gcaaatgggc
      901 gttccattga cgtaaatggg cggtaggcgt gcctaatggg aggtctatat aagcaatgct
      961 cgtttaggga accgccattc tgcctgggga cgtcggagca agcttgattt aggtgacact
     1021 atagaataca agctacttgt tctttttgca ggatccatgc ttcaacttcc tcctcttgaa
     1081 cgtcttactc tcgaggaatt cgccaccatg ggagtgcagg tggaaaccat ctccccagga
     1141 gacgggcgca ccttccccaa gcgcggccag acctgcgtgg tgcactacac cgggatgctt
     1201 gaagatggaa agaaatttga ttcctcccgg gacagaaaca agccctttaa gtttatgcta
     1261 ggcaagcagg aggtgatccg aggctgggaa gaaggggttg cccagatgag tgtgggtcag
     1321 agagccaaac tgactatatc tccagattat gcctatggtg ccactgggca cccaggcatc
     1381 atcccaccac atgccactct cgtcttcgat gtggagcttc taaaactgga aggcagcggc
     1441 gagctcatgg agtttctggc aaccacactg gaacggatcg agaaaaattt cgtgattact
     1501 gatccgagac tgcctgacaa cccaatcatt tttgcgagcg attccttcct gcaactgaca
     1561 gaatattctc gggaagagat cctggggcgc aattgccgtt ttctgcaagg acccgagaca
     1621 gaccgtgcca ctgttcggaa aatcagagat gctattgaca accagactga agtgaccgtt
     1681 cagctgatca attataccaa gagcggcaag aagttctgga acgtgttcca cctgcaaccg
     1741 atgcgcgatt ataagggcga cgtccagtac ttcattggcg tgcagctgga tggcaccgaa
     1801 cgtcttcatg gcgccgctga gcgtgaggcg gtctgcctga tcaaaaagac agcctttcag
     1861 attgctgagg cagcgaacga cgaaaattac tttgcatgct atgaccgagc ccatgttaca
     1921 ggagcatcat caagtagttc ttcaagcacc aaaggcactt acttccctgc aattttgaac
     1981 cctccaccat ccccagccac agagcgatca cattacacta tggaatttgg atattcttca
     2041 aacagtcctt ccactcatag gtcatacagc tacaggccat atagctaccg gcactttgca
     2101 ccccccacca caccctgcag cacagatgtt tgtgacagtg actatgctcc tagtcggaga
     2161 atgacctcag tggcaacagc caagggctat accagtgact tgaactatga ttcagaacct
     2221 gtgcccccac ctcccacacc ccgaagccaa tacttgtcag cagaggagaa ctatgaaagc
     2281 tgcccacctt ctccatacac agagaggagc tattctcatc acctctaccc accgccaccc
     2341 tctccctgta cagactcctc ctagtctaga tagctagcct ccctatagtg agtcgtatta
     2401 cgtagatcca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     2461 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     2521 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     2581 ggaggtgtgg gaggtttttt aattcgcgtc cgcggcgcca atgcattggg cccggtaccc
     2641 agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg
     2701 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cgggagcata
     2761 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     2821 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     2881 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     2941 cgctcggtcg ttcggctgcg ggagcggtat cagctcactc aaaggcggta atacggttat
     3001 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     3061 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     3121 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     3181 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     3241 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     3301 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     3361 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     3421 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     3481 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     3541 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     3601 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     3661 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     3721 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     3781 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     3841 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     3901 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     3961 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     4021 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     4081 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     4141 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     4201 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     4261 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     4321 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     4381 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     4441 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     4501 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     4561 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     4621 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     4681 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     4741 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     4801 aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat
     4861 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     4921 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     4981 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     5041 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     5101 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     5161 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     5221 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     5281 gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg aagggcgatc
     5341 ggtgcgggcc tcttcgctat tacgccagtc gaccgccaat
//
