LOCUS Exported 7147 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Small fragment of split HRP fused to the extracellular terminus of NLG1. CAG promoter.. ACCESSION . VERSION . KEYWORDS CAG sHRPb-NLG SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7147) AUTHORS Martell JD, Yamagata M, Deerinck TJ, Phan S, Kwa CG, Ellisman MH, Sanes JR, Ting AY TITLE A split horseradish peroxidase for the detection of intercellular protein-protein interactions and sensitive visualization of synapses. JOURNAL Nat Biotechnol. 2016 May 30. doi: 10.1038/nbt.3563. PUBMED 27240195 REFERENCE 2 (bases 1 to 7147) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7147 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 4..383 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 386..661 /label=chicken beta-actin promoter intron 662..1670 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 1594..1616 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 1678..1697 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" regulatory 1726..1735 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2179..2220 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" regulatory 2682..2691 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" primer_bind complement(4650..4666) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4650..4666) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4663..4685) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4674..4690 /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(4698..4728) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4743..4764 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 4823..5018 /label=SV40 promoter /note="SV40 early promoter" rep_origin 4869..5004 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4931..4950 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" polyA_signal 5024..5158 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5061..5080) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5115..5134 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5226..5243) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5397..5985) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5477..5496) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6156..7016) /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 6779..6798 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7017..7121) /gene="bla" /label=AmpR promoter ORIGIN 1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata 61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc 121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag 181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac 241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg 301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg 361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc 421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca 481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg 541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt 601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc 661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc 721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc 781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag 841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg 901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg 961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc 1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg 1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc 1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc 1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg 1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct 1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg 1381 gacttccttt gtcccaaatc tggcggagcc gaaatctggg aggcgccgcc gcaccccctc 1441 tagcgggcgc gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt 1501 cgtgcgtcgc cgcgccgccg tccccttctc catctccagc ctcggggctg ccgcaggggg 1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg 1621 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca 1681 acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attccgccac catggcactt 1741 cccagatgca tgtggccaaa ttatgtttgg agagctatga tggcatgtgt ggtccacagg 1801 ggatccggtg ccccattgac tctctgcttg ttgggatgtt tgctacagac ttttcacgta 1861 ctctctcaaa agaccggtaa cttgtctgca ctggttgact ttgacctgcg caccccaacc 1921 atcttcgaca ataagtatta cgtcaatctg gaggaacaga agggcctgat tcagtccgac 1981 caggagctgt tcagttcccc agatgctacc gacactattc ctcttgtgcg cagcttcgct 2041 aatagcacac aaaccttttt caatgctttc gtggaagcta tggatcggat gggaaatata 2101 acgcccctga cgggtacgca gggacagata agacggaact gtagggtcgt gaacagcaac 2161 tctggctcgg gctcgggcgg caagcccatc cccaaccccc tgctgggcct ggacagcacc 2221 caattgttgg atgatgtaga cccattggtt actactaact ttggcaagat taggggaatt 2281 aagaaagaac tcaataatga aattttgggt cctgtcattc agtttcttgg ggttccatat 2341 gccgctccac caacaggaga acatcgtttc cagcctccag aaccaccatc tccctggtct 2401 gacatccgga acgccactca gtttgctcct gtatgtcccc agaatatcat tgatggcaga 2461 ttgcctgaag ttatgcttcc tgtgtggttc actaataact tggatgtggt ttcatcatac 2521 gtccaagacc agagtgaaga ctgtctatac ttaaacatct atgtcccaac tgaagatggt 2581 ccccttacaa agaaacacac agatgattta ggtgataatg acggtgctga agatgaagat 2641 attcgggaca gtgggggtcc caaaccagtg atggtgtaca tccatggcgg ctcttacatg 2701 gaaggtactg gaaatctgta tgatgggagt gtcttggcaa gctatggcaa tgtgatcgtc 2761 atcacagtca actatcggct tggggtactt ggcttcttga gcacagggga tcaggctgcc 2821 aaaggaaact acgggctcct tgacctcatc caggccctaa gatggaccag cgagaacatt 2881 gggttctttg gtggtgaccc cttgcgaatc actgtgtttg gatcaggcgc tgggggttca 2941 tgtgtcaacc tgctgacttt atcccattat tctgaaggta accgttggag caattcaacc 3001 aaaggacttt ttcaacgagc aatagctcag agtggaacag ccctttccag ctgggctgtt 3061 agtttccagc ctgcaaaata cgctagaatt ctggccacaa aagttggctg caatgtttca 3121 gatacagtag agttagtaga atgcctgcag aagaagcctt acaaagaact tgttgatcaa 3181 gatgttcaac cagcccgata ccacatagcc tttggacctg tgatcgatgg tgatgtaata 3241 ccagatgacc ctcagatact gatggaacaa ggagagttcc tcaactatga tataatgttg 3301 ggagttaacc aaggggaagg gttgaagttt gtcgaaaata tagtagatag tgatgatggt 3361 gtatcagcca gtgatttcga ctttgctgtt tctaattttg ttgataattt atatggatat 3421 cctgaaggca aagatgtttt gagagaaacc attaaattca tgtatactga ctgggctgat 3481 cgccataatc ctgaaactag aaggaagaca ttgttggctt tgtttacgga ccatcaatgg 3541 gtagcacctg ctgtggccac agcagacctt cactcgaact ttggctcacc tacatacttc 3601 tatgcctttt atcatcattg ccaaacagat caagttccag cttgggctga tgcagctcat 3661 ggggatgagg ttccctatgt gttgggaatc cccatgattg gccctacaga gttatttcct 3721 tgcaatttct ccaagaatga tgtgatgttg agtgcagtag taatgacata ctggacgaat 3781 tttgctaaaa ctggtgaccc aaatcaacca gttcctcaag acacaaaatt catccatacc 3841 aaacccaacc gctttgaaga agtagcatgg accagatatt cccagaaaga ccagctttat 3901 ctccatattg gattaaaacc gagagttaaa gagcattaca gagccaataa ggtaaatctc 3961 tggctggagc tggtacctca tctgcataat ctcaatgaca tttctcagta tacctcgaca 4021 acaactaaag tgccatccac ggacatcact ctcagaccta caaggaagaa ttccacacca 4081 gtcacatcag cctttcccac tgccaaacag gatgatccca agcaacaacc aagccccttc 4141 tcggtggatc agagggacta ctccacagag ctaagtgtca ctatcgcagt gggggcctct 4201 ctgctgtttc tcaacatctt ggcttttgca gccctgtact acaagaagga taagaggaga 4261 catgatgtcc accggaggtg cagccctcag cgcacgacca ccaacgacct aacccatgct 4321 ccagaagagg aaattatgtc tctccaaatg aagcacactg acttggatca cgagtgtgag 4381 tccatccatc cacatgaggt ggttcttcgg accgcctgtc ccccagatta tactctagct 4441 atgaggaggt cacctgatga tattccacta atgacaccta acaccatcac aatgattccc 4501 aacactatac cagggattca gcccttacat acattcaaca catttactgg aggacagaat 4561 aatacactgc cccatcccca cccacacccc cattcacatt caacaaccag ggtatagaga 4621 tctgcggccg caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat 4681 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc 4741 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga 4801 aacctgtcgt gccagcggat ccgcatctca attagtcagc aaccatagtc ccgcccctaa 4861 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac 4921 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt 4981 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctaacttgt ttattgcagc 5041 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc 5101 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatccg 5161 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc 5221 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct 5281 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg 5341 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc 5401 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga 5461 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct 5521 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg 5581 gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag 5641 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat 5701 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac 5761 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac 5821 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc 5881 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt 5941 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc 6001 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg 6061 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca 6121 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca 6181 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag 6241 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac 6301 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc 6361 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct 6421 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc 6481 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg 6541 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc 6601 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat 6661 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag 6721 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat 6781 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg 6841 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca 6901 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga 6961 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc 7021 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata 7081 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg 7141 ccacctg //