LOCUS       sequence-384904-        6632 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human beta-actin labelled with near-infrared fluorescent protein 
            miRFP718nano.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6632)
  AUTHORS   Oliinyk OS, Ma C, Pletnev S, Baloban M, Toboada C, Sheng H, Yao J, 
            Verkhusha VV
  TITLE     Deep-tissue SWIR imaging using rationally designed small red-shifted
            near-infrared fluorescent protein.
  JOURNAL   Nat Methods. 2022 Dec 1. doi: 10.1038/s41592-022-01683-0.
  PUBMED    36456785
REFERENCE   2  (bases 1 to 6632)
  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 
            Methods. 2022 Dec 1. doi: 10.1038/s41592-022-01683-0."
COMMENT     Sequence Label: pActin-miRFP718nano
FEATURES             Location/Qualifiers
     source          1..6632
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     111..130
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    263..487
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     476..493
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(516..533)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(549..578)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(2251..2270)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2252..2270)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2315..2518)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2344..2364)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2519..2898
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     3070..3089
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3164..3268
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3269..3337,3338..4129)
                     /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(3487..3506)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4300..4888
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4789..4808
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5042..5059
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5176..5197
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5212..5217,5218..5235,5236..5242)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5250..5266
                     /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)."
     primer_bind     5255..5277
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5274..5290
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5274..5290
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    5339..5460
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5350..5369)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5404..5423
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5590..5964)
                     /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"
     promoter        complement(5983..6030)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(6078..6407)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6092..6227
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6146..6165)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     6392..6412
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(join(6421..6632,1..217))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6531..6552)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
       61 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
      121 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
      181 acacccgccg cgcttaatgc gccgctacag ggcgcgtggg gataccccct agagccccag
      241 ctggttcttt ccgcctcaga agccatagag cccaccgcat ccccagcatg cctgctattg
      301 tcttcccaat cctccccctt gctgtcctgc cccaccccac cccccagaat agaatgacac
      361 ctactcagac aatgcgatgc aatttcctca ttttattagg aaaggacagt gggagtggca
      421 ccttccaggg tcaaggaagg cacgggggag gggcaaacaa cagatggctg gcaactagaa
      481 ggcacagtcg aggctgatca gcgggtttaa actcaatggt gatggtgatg atgaccggta
      541 tgcatattca gatcctcttc tgagatgagt ttttgttcga agggccctct agactagaag
      601 catttgcggt ggacgatgga ggggccggac tcgtcatact cctgcttgct gatccacatc
      661 tgctggaagg tggacagcga ggccaggatg gagccgccga tccacacgga gtacttgcgc
      721 tcaggaggag caatgatctt gatcttcatt gtgctgggtg ccagggcagt gatctccttc
      781 tgcatcctgt cggcaatgcc agggtacatg gtggtgccgc cagacagcac tgtgttggcg
      841 tacaggtctt tgcggatgtc cacgtcacac ttcatgatgg agttgaaggt agtttcgtgg
      901 atgccacagg actccatgcc caggaaggaa ggctggaaga gtgcctcagg gcagcggaac
      961 cgctcattgc caatggtgat gacctggccg tcaggcagct cgtagctctt ctccagggag
     1021 gagctggaag cagccgtggc catctcttgc tcgaagtcca gggcgacgta gcacagcttc
     1081 tccttaatgt cacgcacgat ttcccgctcg gccgtggtgg tgaagctgta gccgcgctcg
     1141 gtgaggatct tcatgaggta gtcagtcagg tcccggccag ccaggtccag acgcaggatg
     1201 gcatggggga gggcataccc ctcgtagatg ggcacagtgt gggtgacccc gtcaccggag
     1261 tccatcacga tgccagtggt acggccagag gcgtacaggg atagcacagc ctggatagca
     1321 acgtacatgg ctggggtgtt gaaggtctca aacatgatct gggtcatctt ctcgcggttg
     1381 gccttggggt tcaggggggc ctcggtcagc agcacggggt gctcctcggg agccacacgc
     1441 agctcattgt agaaggtgtg gtgccagatt ttctccatgt cgtcccagtt ggtgacgatg
     1501 ccgtgctcga tggggtactt cagggtgagg atgcctctct tgctctgggc ctcgtcgccc
     1561 acataggaat ccttctgacc catgcccacc atcacgccct ggtgcctggg gcgccccacg
     1621 atggagggga agacggcccg gggggcatcg tcgcccgcga agccggcctt gcacatgccg
     1681 gagccgttgt cgacgacgag cgcggcgata tcatcatcca tagatcttcc tcctgatcca
     1741 ccacctccag aaccacctcc tcctgatcca ccacctccgc tctgctggat ggcgatgccc
     1801 atcaccacgg cttgctgttt caggaactcg atttcccagg cctgccagct ccgaggggcg
     1861 gccagttggt gggcgctgaa caagccccac agcttcttgc ccttcaggat ggggacagcc
     1921 acaatggccc gcacctggta ctgggtcagg aagtctctat agctctccgc cagattggcc
     1981 tggtggatgt cggcgatggc ctggtatctg ccgtgcaggt attcctcgcc tctggtttcc
     2041 atgaagtagc agtcccgcac gtgggtcttc aggatgctca accatccgtc gtccacggct
     2101 tcgtagatga tgttgccgga gtaatcttcc tcgaacttga acacgcagag tctgtccact
     2161 tgcaggaact ggcgcacctc ggtcacgatg gtgttcagca tcttgtccag gtttgccatg
     2221 gtaccaagct taactagcca gcttgggtct ccctatagtg agtcgtatta atttcgataa
     2281 gccagtaagc agtgggttct ctagttagcc agagagctct gcttatatag acctcccacc
     2341 gtacacgcct accgcccatt tgcgtcaatg gggcggagtt gttacgacat tttggaaagt
     2401 cccgttgatt ttggtgccaa aacaaactcc cattgacgtc aatggggtgg agacttggaa
     2461 atccccgtga gtcaaaccgc tatccacgcc cattgatgta ctgccaaaac cgcatcacca
     2521 tggtaatagc gatgactaat acgtagatgt actgccaagt aggaaagtcc cataaggtca
     2581 tgtactgggc ataatgccag gcgggccatt taccgtcatt gacgtcaata gggggcgtac
     2641 ttggcatatg atacacttga tgtactgcca agtgggcagt ttaccgtaaa tactccaccc
     2701 attgacgtca atggaaagtc cctattggcg ttactatggg aacatacgtc attattgacg
     2761 tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc atttaccgta agttatgtaa
     2821 cgcggaactc catatatggg ctatgaacta atgaccccgt aattgattac tattaataac
     2881 tagtcaataa tcaatgtcaa cgcgtatatc tggcccgtac atcgcgaagc agcgcaaaac
     2941 gcctaaccct aagcagattc ttcatgcaat tgtcggtcaa gccttgcctt gttgtagctt
     3001 aaattttgct cgcgcactac tcagcgacct ccaacacaca agcagggagc agatactggc
     3061 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatagggga tcgggagatc
     3121 tcccgatccg tcgacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     3181 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     3241 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     3301 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     3361 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     3421 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     3481 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     3541 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     3601 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     3661 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     3721 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     3781 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     3841 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     3901 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     3961 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     4021 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     4081 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     4141 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     4201 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     4261 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     4321 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     4381 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     4441 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     4501 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     4561 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     4621 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     4681 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     4741 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     4801 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     4861 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     4921 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     4981 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     5041 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     5101 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     5161 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     5221 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     5281 cagctatgac catgattacg ccaagctcta gctagaggtc gacggtatac agacatgata
     5341 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     5401 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     5461 ggggtgggcg aagaactcca gcatgagatc cccgcgctgg aggatcatcc agccggcgtc
     5521 ccggaaaacg attccgaagc ccaacctttc atagaaggcg gcggtggaat cgaaatctcg
     5581 tagcacgtgt cagtcctgct cctcggccac gaagtgcacg cagttgccgg ccgggtcgcg
     5641 cagggcgaac tcccgccccc acggctgctc gccgatctcg gtcatggccg gcccggaggc
     5701 gtcccggaag ttcgtggaca cgacctccga ccactcggcg tacagctcgt ccaggccgcg
     5761 cacccacacc caggccaggg tgttgtccgg caccacctgg tcctggaccg cgctgatgaa
     5821 cagggtcacg tcgtcccgga ccacaccggc gaagtcgtcc tccacgaagt cccgggagaa
     5881 cccgagccgg tcggtccaga actcgaccgc tccggcgacg tcgcgcgcgg tgagcaccgg
     5941 aacggcactg gtcaacttgg ccatggttta gttcctcacc ttgtcgtatt atactatgcc
     6001 gatatactat gccgatgatt aattgtcaac acgtgctgat cagatccgaa aatggatata
     6061 caagctcccg ggagcttttt gcaaaagcct aggcctccaa aaaagcctcc tcactacttc
     6121 tggaatagct cagaggcaga ggcggcctcg gcctctgcat aaataaaaaa aattagtcag
     6181 ccatggggcg gagaatgggc ggaactgggc ggagttaggg gcgggatggg cggagttagg
     6241 ggcgggacta tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg
     6301 gagcctgggg actttccaca cctggttgct gactaattga gatgcatgct ttgcatactt
     6361 ctgcctgctg gggagcctgg ggactttcca caccctaact gacacacatt ccacagaatt
     6421 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     6481 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     6541 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     6601 agggcgatgg cccactacgt gaaccatcac cc
//
