LOCUS       Exported                5899 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  I-kappa-B-alpha fused with near-infrared fluorescent protein 
            miRFP703 (I-kappa-B-alpha reporter).
ACCESSION   .
VERSION     .
KEYWORDS    pI-kappa-B-alpha-miRFP703
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5899)
  AUTHORS   Shcherbakova DM, Baloban M, Emelyanov AV, Brenowitz M, Guo P, 
            Verkhusha VV
  TITLE     Bright monomeric near-infrared fluorescent proteins as tags and 
            biosensors for multiscale imaging.
  JOURNAL   Nat Commun. 2016 Aug 19;7:12405. doi: 10.1038/ncomms12405.
  PUBMED    27539380
REFERENCE   2  (bases 1 to 5899)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5899
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             1..948
                     /codon_start=1
                     /product="bright monomeric near-infrared fluorescent 
                     protein engineered from a bacterial phytochrome, with an 
                     emission maximum of 703 nm (Shcherbakova et al., 2016)"
                     /label=miRFP703
                     /label=ORF frame 1
                     /translation="MVAGHASGSPAFGTASHSNCEHEEIHLAGSIQPHGALLVVSEHDH
                     RVIQASANAAEFLNLGSVLGVPLAEIDGDLLIKILPHLDPTAEGMPVAVRCRIGNPSTE
                     YCGLMHRPPEGGLIIELERAGPSIDLSGTLAPALERIRTAGSLRALCDDTVLLFQQCTG
                     YDRVMVYRFDEQGHGLVFSECHVPGLESYFGNRYPSSLVPQMARQLYVRQRVRVLVDVT
                     YQPVPLEPRLSPLTGRDLDMSGCFLRSMSPIHLQFLKDMGVRATLAVSLVVGGKLWGLV
                     VCHHYLPRFIRFELRAICKRLAERIATRITALES"
     polyA_signal    1070..1191
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1107..1126)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1161..1180
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1198..1653)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1335..1356)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1547..1566
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1680..1784
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1786..2143
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(1809..1829)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1994..2129
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2056..2075
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2178..2972
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2232..2251)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2842..2861
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3160..3179)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3204..3251
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3580..4168
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4069..4088
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4343..4646
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4647..4850
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4801..4821
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 atggtagcag gtcatgcctc tggcagcccc gcattcggga ccgcctctca ttcgaattgc
       61 gaacatgaag agatccacct cgccggctcg atccagccgc atggcgcgct tctggtcgtc
      121 agcgaacatg atcatcgcgt catccaggcc agcgccaacg ccgcggaatt tctgaatctc
      181 ggaagcgtac tcggcgttcc gctcgccgag atcgacggcg atctgttgat caagatcctg
      241 ccgcatctcg atcccaccgc cgaaggcatg ccggtcgcgg tgcgctgccg gatcggcaat
      301 ccctctacgg agtactgcgg tctgatgcat cggcctccgg aaggcgggct gatcatcgaa
      361 ctcgaacgtg ccggcccgtc gatcgatctg tcaggcacgc tggcgccggc gctggagcgg
      421 atccgcacgg cgggttcact gcgcgcgctg tgcgatgaca ccgtgctgct gtttcagcag
      481 tgcaccggct acgaccgggt gatggtgtat cgtttcgatg agcaaggcca cggcctggta
      541 ttctccgagt gccatgtgcc tgggctcgaa tcctatttcg gcaaccgcta tccgtcgtcg
      601 ctggtcccgc agatggcgcg gcagctgtac gtgcggcagc gcgtccgcgt gctggtcgac
      661 gtcacctatc agccggtgcc gctggagccg cggctatcgc cgctgaccgg gcgcgatctc
      721 gacatgtcgg gctgcttcct gcgctcgatg tcgccgatcc atctgcagtt cctgaaggac
      781 atgggcgtgc gcgccaccct ggcggtgtcg ctggtggtcg gcggcaagct gtggggcctg
      841 gttgtctgtc accattatct gccgcgcttc atccgtttcg agctgcgggc gatctgcaaa
      901 cggctcgccg aaaggatcgc gacgcggatc accgcgcttg agagctaagc ggccgcgact
      961 ctagatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
     1021 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     1081 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     1141 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa
     1201 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     1261 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     1321 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     1381 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     1441 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     1501 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     1561 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     1621 ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc
     1681 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     1741 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa
     1801 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     1861 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     1921 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     1981 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     2041 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     2101 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag
     2161 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     2221 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     2281 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     2341 gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     2401 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     2461 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     2521 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     2581 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     2641 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     2701 aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     2761 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     2821 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     2881 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     2941 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     3001 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     3061 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     3121 catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat
     3181 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg
     3241 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc
     3301 gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag
     3361 ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc
     3421 aggttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     3481 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     3541 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     3601 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     3661 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     3721 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     3781 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     3841 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     3901 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     3961 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     4021 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     4081 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     4141 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     4201 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     4261 taaccgtatt accgccatgc attagttatt aatagtaatc aattacgggg tcattagttc
     4321 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
     4381 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     4441 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     4501 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
     4561 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
     4621 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
     4681 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
     4741 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
     4801 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga
     4861 accgtcagat ccgctagcgc taccggactc agatctcatg ttccaggcgg ccgagcgtcc
     4921 ccaggagtgg gccatggagg gcccccgcga cgggctgaag aaggagcggc tactggacga
     4981 ccgccacgac agcggcctgg actccatgaa agacgaggag tacgagcaga tggtcaagga
     5041 gctgcaggag atccgcctcg agccgcagga ggtgccgcgc ggctcggagc cctggaagca
     5101 gcagctcacc gaggacgggg actcgttcct gcacttggcc atcatccatg aagaaaaggc
     5161 actgaccatg gaagtgatcc gccaggtgaa gggagacctg gccttcctca acttccagaa
     5221 caacctgcag cagactccac tccacttggc tgtgatcacc aaccagccag aaattgctga
     5281 ggcacttctg ggagctggct gtgatcctga gctccgagac tttcgaggaa atacccccct
     5341 acaccttgcc tgtgagcagg gctgcctggc cagcgtggga gtcctgactc agtcctgcac
     5401 caccccgcac ctccactcca tcctgaaggc taccaactac aatggccaca cgtgtctaca
     5461 cttagcctct atccatggct acctgggcat cgtggagctt ttggtgtcct tgggtgctga
     5521 tgtcaatgct caggagccct gtaatggccg gactgccctt cacctcgcag tggacctgca
     5581 aaatcctgac ctggtgtcac tcctgttgaa gtgtggggct gatgtcaaca gagttaccta
     5641 ccagggctat tctccctacc agctcacctg gggccgccca agcacccgga tacagcagca
     5701 gctgggccag ctgacactag aaaaccttca gatgctgcca gagagtgagg atgaggagag
     5761 ctatgacaca gagtcagagt tcacggagtt cacagaggac gagctgccct atgatgactg
     5821 tgtgtttgga ggccagcgtc tgacgttagg ttcctcggga ggttccagtt ctggaggggg
     5881 ttccggaggt ggtggatca
//
