LOCUS Exported 4797 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Constitutive expression of Antares in bacteria. ACCESSION . VERSION . KEYWORDS pNCS-Antares SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4797) AUTHORS Chu J, Oh Y, Sens A, Ataie N, Dana H, Macklin JJ, Laviv T, Welf ES, Dean KM, Zhang F, Kim BB, Tang CT, Hu M, Baird MA, Davidson MW, Kay MA, Fiolka R, Yasuda R, Kim DS, Ng HL, Lin MZ TITLE A bright cyan-excitable orange fluorescent protein facilitates dual-emission microscopy and enhances bioluminescence imaging in vivo. JOURNAL Nat Biotechnol. 2016 May 30. doi: 10.1038/nbt.3550. PUBMED 27240196 REFERENCE 2 (bases 1 to 4797) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4797 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 20..39 /label=T7 /note="T7 promoter, forward primer" promoter 20..38 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 164..181 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 185..217 /codon_start=1 /product="leader peptide from bacteriophage T7 gene 10" /label=T7 tag (gene 10 leader) /note="promotes efficient translation in E. coli" /translation="MASMTGGQQMG" CDS 221..244 /codon_start=1 /product="Xpress(TM) epitope tag, including an enterokinase recognition and cleavage site" /label=Xpress(TM) tag /translation="DLYDDDDK" CDS 935..1441 /codon_start=1 /product="NanoLuc(R) luciferase" /label=Nluc /note="human codon-optimized" /translation="FTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRIV LSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVTP NMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWRL CERILA" primer_bind complement(2198..2216) /label=T7 Term /note="T7 terminator, reverse primer" terminator 2212..2259 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin 2356..2811 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2443..2462) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2653..2674 /label=F1ori-F /note="F1 origin, forward primer" promoter 2837..2941 /gene="bla" /label=AmpR promoter CDS 2942..3802 /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 complement(3160..3179) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 3973..4561 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4462..4481 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4715..4732 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct 61 agaatcaccg agtttattct tgacacctga tgcgatgaat gatataatag gaaagtactg 121 ttttgtttaa ctttaagaag gggatataca tatgcggggt tctcatcatc atcatcatca 181 tggtatggct agcatgactg gtggacagca aatgggtcgg gatctgtacg acgatgacga 241 taaggatccg atggtgagca agggcgagga gctgatcaag gagaacatga gaagcaagct 301 gtacctggaa ggcagcgtga acggccacca gttcaagtgc acccacgaag gggagggcaa 361 gccctacgag ggcaagcaga ccaacaggat caaggtggtg gagggaggcc ccctgccgtt 421 cgcattcgac atcctggcca cccactttat gtacgggagc aaggtgttca tcaagtaccc 481 cgccgacctc cccgattatt ttaagcagtc cttccctgag ggcttcacat gggagagagt 541 catggtgttc gaagacgggg gcgtgctgac cgccacccag gacaccagcc tccaggacgg 601 cgagctcatc tacaacgtca aggtcagagg ggtgaacttc ccagccaacg gccccgtgat 661 gcagaagaaa acactgggct gggagcccag caccgagacc atgtaccccg ctgacggcgg 721 cctggaaggc agatgcgaca aggccctgaa gctcgtgggc gggggccacc tgcacgtcaa 781 cttcaagacc acatacaagt ccaagaaacc cgtgaagatg cccggcgtcc actacgtgga 841 ccgcagactg gaaagaatca aggaggccga caacgagacc tacgtcgagc agtacgagca 901 cgctgtggcc agatactcca acctgggcgg aggcttcaca ctcgaagatt tcgttgggga 961 ctggcgacag acagccggct acaacctgga ccaagtcctt gaacagggag gtgtgtccag 1021 tttgtttcag aatctcgggg tgtccgtaac tccgatccaa aggattgtcc tgagcggtga 1081 aaatgggctg aagatcgaca tccatgtcat catcccgtat gaaggtctga gcggcgacca 1141 aatgggccag atcgaaaaaa tttttaaggt ggtgtaccct gtggatgatc atcactttaa 1201 ggtgatcctg cactatggca cactggtaat cgacggggtt acgccgaaca tgatcgacta 1261 tttcggacgg ccgtatgaag gcatcgccgt gttcgacggc aaaaagatca ctgtaacagg 1321 gaccctgtgg aacggcaaca aaattatcga cgagcgcctg atcaaccccg acggctccct 1381 gctgttccga gtaaccatca acggagtgac cggctggcgg ctgtgcgaac gcattctggc 1441 gaggcacgag ctgatcaagg agaacatgag aagcaagctg tacctggaag gcagcgtgaa 1501 cggccaccag ttcaagtgca cccacgaagg ggagggcaag ccctacgagg gcaagcagac 1561 caacaggatc aaggtggtgg agggaggccc cctgccgttc gcattcgaca tcctggccac 1621 ccactttatg tacgggagca aggtgttcat caagtacccc gccgacctcc ccgattattt 1681 taagcagtcc ttccctgagg gcttcacatg ggagagagtc atggtgttcg aagacggggg 1741 cgtgctgacc gccacccagg acaccagcct ccaggacggc gagctcatct acaacgtcaa 1801 ggtcagaggg gtgaacttcc cagccaacgg ccccgtgatg cagaagaaaa cactgggctg 1861 ggagcccagc accgagacca tgtaccccgc tgacggcggc ctggaaggca gatgcgacaa 1921 ggccctgaag ctcgtgggcg ggggccacct gcacgtcaac ttcaagacca catacaagtc 1981 caagaaaccc gtgaagatgc ccggcgtcca ctacgtggac cgcagactgg aaagaatcaa 2041 ggaggccgac aacgagacct acgtcgagca gtacgagcac gctgtggcca gatactccaa 2101 cctgggcgga ggcatggacg agctgtacaa gtaatgaatt cgaagcttga tccggctgct 2161 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa 2221 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc 2281 ggatctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc 2341 tgaatggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta 2401 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc 2461 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt 2521 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg 2581 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca 2641 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct 2701 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga 2761 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac 2821 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat 2881 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag 2941 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc 3001 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc 3061 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc 3121 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc 3181 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt 3241 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt 3301 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat 3361 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct 3421 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat 3481 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc 3541 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg 3601 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc 3661 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta 3721 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc 3781 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga 3841 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat 3901 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat 3961 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa 4021 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa 4081 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt 4141 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt 4201 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata 4261 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt 4321 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac 4381 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga 4441 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg 4501 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa 4561 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat 4621 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc 4681 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga 4741 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcag //