LOCUS Exported 5352 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses 3xFNLDD in mammalian cells. ACCESSION . VERSION . KEYWORDS 3xFNLDD/pCMV-7.1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5352) AUTHORS Fujita T, Kitaura F, Fujii H TITLE A Critical Role of the Thy28-MYH9 Axis in B Cell-Specific Expression of the Pax5 Gene in Chicken B Cells. JOURNAL PLoS One. 2015 Jan 21;10(1):e0116579. doi: 10.1371/journal.pone.0116579. eCollection 2015. PUBMED 25607658 REFERENCE 2 (bases 1 to 5352) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5352 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 126..148 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 140..157 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 141..157 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" enhancer 318..697 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 698..901 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 852..872 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" CDS 931..996 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" CDS 1033..1053 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 1066..1674 /codon_start=1 /gene="lexA" /product="E. coli DNA-binding protein that represses genes involved in the SOS response to DNA damage" /label=LexA /note="The N-terminal DNA-binding domain recognizes the 16-bp palindromic sequence 5'-CTGTATATATATACAG-3'." /translation="MKALTARQQEVFDLIRDHISQTGMPPTRAEIAQRLGFRSPNAAEE HLKALARKGVIEIVSGASRGIRLLQEEEEGLPLVGRVAAGEPLLAQQHIEGHYQVDPSL FKPNADFLLRVSGMSMKDIGIMDGDLLAVHKTQDVRNGQVVVARIDDEVTVKRLKKQGN KVELLPENSEFKPIVVDLRQQSFTIEGLAVGVIRNGDWL" primer_bind 1609..1630 /label=LexA /note="LexA DNA binding domain, forward primer" polyA_signal 1693..2315 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(1892..1911) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" primer_bind complement(2339..2359) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2344..2673 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 2524..2659 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2586..2605 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(2711..2730) /label=T7 /note="T7 promoter, forward primer" promoter complement(2712..2730) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2744..2760) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2744..2760) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2757..2779) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2768..2784 /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)." promoter complement(2792..2822) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2837..2858 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2975..2992) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3146..3734) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3226..3245) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3905..4765) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 4528..4547 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4766..4870) /gene="bla" /label=AmpR promoter rep_origin 4897..5352 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4984..5003) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5194..5215 /label=F1ori-F /note="F1 origin, forward primer" ORIGIN 1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct 61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg 121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa 181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta 241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc 301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg 361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc 421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat 481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc 541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga 601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg 661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac 721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt 781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc 841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc 901 tcgtttagtg aaccgtcaga attaaccatg gactacaaag accatgacgg tgattataaa 961 gatcatgaca tcgattacaa ggatgacgat gacaagcttg cggccgcgaa ttcatcgata 1021 gatcgatccc ctccaaaaaa gaagagaaag gtcgaacctg agggaatgaa agcgttaacg 1081 gccaggcaac aagaggtgtt tgatctcatc cgtgatcaca tcagccagac aggtatgccg 1141 ccgacgcgtg cggaaatcgc gcagcgtttg gggttccgtt ccccaaacgc ggctgaagaa 1201 catctgaagg cgctggcacg caaaggcgtt attgaaattg tttccggcgc atcacgcggg 1261 attcgtctgt tgcaggaaga ggaagaaggg ttgccgctgg taggtcgtgt ggctgccggt 1321 gaaccacttc tggcgcaaca gcatattgaa ggtcattatc aggtcgatcc ttccttattc 1381 aagccgaatg ctgatttcct gctgcgcgtc agcgggatgt cgatgaaaga tatcggcatt 1441 atggatggtg acttgctggc agtgcataaa actcaggatg tacgtaacgg tcaggtcgtt 1501 gtcgcacgta ttgatgacga ggttaccgtt aagcgcctga aaaaacaggg caataaagtc 1561 gaactgttgc cagaaaatag cgagtttaaa ccaattgtcg tagatcttcg tcagcagagc 1621 ttcaccattg aagggctggc ggttggggtt attcgcaacg gcgactggct gtagtcgact 1681 ctagaggatc ccgggtggca tccctgtgac ccctccccag tgcctctcct ggccctggaa 1741 gttgccactc cagtgcccac cagccttgtc ctaataaaat taagttgcat cattttgtct 1801 gactaggtgt ccttctataa tattatgggg tggagggggg tggtatggag caaggggcaa 1861 gttgggaaga caacctgtag ggcctgcggg gtctattggg aaccaagctg gagtgcagtg 1921 gcacaatctt ggctcactgc aatctccgcc tcctgggttc aagcgattct cctgcctcag 1981 cctcccgagt tgttgggatt ccaggcatgc atgaccaggc tcagctaatt tttgtttttt 2041 tggtagagac ggggtttcac catattggcc aggctggtct ccaactccta atctcaggtg 2101 atctacccac cttggcctcc caaattgctg ggattacagg cgtgaaccac tgctcccttc 2161 cctgtccttc tgattttaaa ataactatac cagcaggagg acgtccagac acagcatagg 2221 ctacctggcc atgcccaacc ggtgggacat ttgagttgct tgcttggcac tgtcctctca 2281 tgcgttgggt ccactcagta gatgcctgtt gaattgggta cgcggccagc ttggctgtgg 2341 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa 2401 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc 2461 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg 2521 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt 2581 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga 2641 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc tcgaggaact gaaaaaccag 2701 aaagttaatt ccctatagtg agtcgtatta aattcgtaat catgtcatag ctgtttcctg 2761 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta 2821 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg 2881 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga 2941 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg 3001 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag 3061 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc 3121 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca 3181 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt 3241 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc 3301 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc 3361 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc 3421 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact 3481 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg 3541 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta 3601 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca 3661 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa 3721 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg 3781 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc 3841 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg 3901 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat 3961 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg 4021 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa 4081 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca 4141 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc 4201 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt 4261 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa 4321 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat 4381 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct 4441 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga 4501 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag 4561 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga 4621 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca 4681 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg 4741 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc 4801 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag 4861 gggttccgcg cacatttccc cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa 4921 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc 4981 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag 5041 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca 5101 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc 5161 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa 5221 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct 5281 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa 5341 cgcttacaat tt //