LOCUS       sequence-139801-        4797 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression of Antares in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
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 Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Biotechnol. 2016 May 30. doi: 10.1038/nbt.3550."
COMMENT     Sequence Label: pNCS-Antares
FEATURES             Location/Qualifiers
     source          1..4797
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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             join(221..229,230..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             join(2942..3010,3011..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
//
