LOCUS       sequence-318341-        5491 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Monomeric near-infrared fluorescent protein miRFP670nano in 
            mammalian plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5491)
  AUTHORS   Oliinyk OS, Shemetov AA, Pletnev S, Shcherbakova DM, Verkhusha VV
  TITLE     Smallest near-infrared fluorescent protein evolved from 
            cyanobacteriochrome as versatile tag for spectral multiplexing.
  JOURNAL   Nat Commun. 2019 Jan 17;10(1):279. doi: 10.1038/s41467-018-08050-8.
  PUBMED    30655515
REFERENCE   2  (bases 1 to 5491)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2019-01-17"; volume: "10(1):279. doi"; pages: " 
            10.1038/s41467-018-08050-8"
COMMENT     Sequence Label: miRFP670nano-pcDNA
FEATURES             Location/Qualifiers
     source          1..5491
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        152..531
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        532..735
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     686..706
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     780..799
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        780..798
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1331..1360
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1376..1393
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1416..1433)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1422..1646
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1692..2120
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1779..1798)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1989..2010
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2129..2149)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2134..2463
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2314..2449
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2376..2395
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2511..2558
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2577..2951
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    3081..3202
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3118..3137)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3172..3191
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3251..3267)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3251..3267)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3264..3286)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3275..3291
                     /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(join(3299..3305,3306..3323,3324..3329))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3344..3365
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3482..3499)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3653..4241)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3733..3752)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4412..5203,5204..5272))
                     /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     5035..5054
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5273..5377)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5452..5471)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca
       61 aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg
      121 cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag
      181 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
      241 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
      301 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
      361 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
      421 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
      481 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
      541 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
      601 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
      661 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
      721 tatataagca gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt
      781 aatacgactc actataggga gacccaagct ggctagttaa gcttggtacc atggcaaacc
      841 tggacaagat gctgaatacc acagtaacag aggtgcggca gttcctgcag gtggacagag
      901 tgtgcgtgtt ccagtttgag gaggattata gcggagtggt ggtggtggag gccgtggacg
      961 ataggtggat ctccatcctg aagacccagg tgcgggatag atacttcatg gagacaaggg
     1021 gcgaggagta ttctcacggc cgctaccagg ccatcgccga catctacacc gcaaacctga
     1081 cagagtgcta cagggatctg ctgacacagt ttcaggtgag agcaatcctg gccgtgccca
     1141 tcctgcaggg caagaagctg tggggcctgt tggtggcaca ccagctggcg gcccctagac
     1201 agtggcagac ctgggagatc gactttctga agcagcaggc cgtggtggtg ggcatcgcca
     1261 tccagcagag ctagctgaat tctgcagata tccagcacag tggcggccgc tcgagtctag
     1321 agggcccttc gaacaaaaac tcatctcaga agaggatctg aatatgcata ccggtcatca
     1381 tcaccatcac cattgagttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     1441 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     1501 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     1561 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     1621 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     1681 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     1741 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     1801 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     1861 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     1921 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     1981 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     2041 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     2101 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     2161 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     2221 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     2281 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     2341 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     2401 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     2461 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcagcacgt gttgacaatt
     2521 aatcatcggc atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaaccatgg
     2581 ccaagttgac cagtgccgtt ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt
     2641 tctggaccga ccggctcggg ttctcccggg acttcgtgga ggacgacttc gccggtgtgg
     2701 tccgggacga cgtgaccctg ttcatcagcg cggtccagga ccaggtggtg ccggacaaca
     2761 ccctggcctg ggtgtgggtg cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg
     2821 tgtccacgaa cttccgggac gcctccgggc cggccatgac cgagatcggc gagcagccgt
     2881 gggggcggga gttcgccctg cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg
     2941 agcaggactg acacgtgcta cgagatttcg attccaccgc cgccttctat gaaaggttgg
     3001 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     3061 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     3121 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     3181 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     3241 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     3301 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     3361 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     3421 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     3481 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     3541 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     3601 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     3661 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     3721 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     3781 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     3841 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     3901 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     3961 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     4021 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     4081 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     4141 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     4201 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     4261 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     4321 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     4381 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     4441 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     4501 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     4561 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     4621 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     4681 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     4741 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     4801 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     4861 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     4921 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     4981 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     5041 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     5101 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     5161 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     5221 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     5281 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     5341 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     5401 ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg
     5461 ctctgatgcc gcatagttaa gccagtatct g
//
