LOCUS       sequence-365038-        5510 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression, miRFP670nano3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5510)
  AUTHORS   Oliinyk OS, Baloban M, Clark CL, Carey E, Pletnev S, Nimmerjahn A, 
            Verkhusha VV
  TITLE     Single-domain near-infrared protein provides a scaffold for 
            antigen-dependent fluorescent nanobodies.
  JOURNAL   Nat Methods. 2022 May 23. pii: 10.1038/s41592-022-01467-6. doi: 
            10.1038/s41592-022-01467-6.
  PUBMED    35606446
REFERENCE   2  (bases 1 to 5510)
  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 May 23. pii: 10.1038/s41592-022-01467-6. doi: 
            10.1038/s41592-022-01467-6."
COMMENT     Sequence Label: pAAV-hSYN-miRFP670nano3-WPRE-hGHpA
FEATURES             Location/Qualifiers
     source          1..5510
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        266..569
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        592..1039
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      1163..1172
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    1637..2225
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1690..1710)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2108..2119)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2257..2733
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2456..2475)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2773..2913
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2773..2902
                     /label=AAV2 ITR
     rep_origin      2988..3443
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3075..3094)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3285..3306
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3460..3479)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3579..3601
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3639..3657)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3725..3829
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3830..3898,3899..4690)
                     /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(4048..4067)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4861..5449
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5350..5369
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtgcggcg acgatagtcg aagttcctat actctagtaa
      181 gaataggaac ttcggatctc ggccgtagtt attaatagta atcaattacg gggtcattag
      241 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      301 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      361 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      421 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      481 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      541 tctacgtatt agtcatcgct attaccatgg ctgcagaggg ccctgcgtat gagtgcaagt
      601 gggttttagg accaggatga ggcggggtgg gggtgcctac ctgacgaccg accccgaccc
      661 actggacaag cacccaaccc ccattcccca aattgcgcat cccctatcag agagggggag
      721 gggaaacagg atgcggcgag gcgcgtgcgc actgccagct tcagcaccgc ggacagtgcc
      781 ttcgcccccg cctggcggcg cgcgccaccg ccgcctcagc actgaaggcg cgctgacgtc
      841 actcgccggt cccccgcaaa ctccccttcc cggccacctt ggtcgcgtcc gcgccgccgc
      901 cggcccagcc ggaccgcacc acgcgaggcg cgagataggg gggcacgggc gcgaccatct
      961 gcgctgcggc gccggcgact cagcgctgcc tcagtctgcg gtgggcagcg gaggagtcgt
     1021 gtcgtgcctg agagcgcagt cgagttaacg gtaccggccc cgggagacgg cggcggtggc
     1081 ggcgcgggca gagcaaggac gcggcggatc ccactcgcac agcagcgcac tcggtgcccc
     1141 gcgcagggtc ggtaccgaat tcgccaccat ggcaaacctg gacaagatgc tgaacaccac
     1201 cgtgaccgag gtgcgcaagt tcctgcaagc ggacagagtg tgcgtgttca agttcgagga
     1261 agattactcc ggcaccgtct cgcacgaagc cgtggacgac agatggatta gcatcctgaa
     1321 gacccaggtg caggacagat acttcatgga aaccagaggc gaggaatacg tccacggcag
     1381 ataccaggcc atcgccgaca tctacacagc caatctggtc gagtgctaca gagacctgct
     1441 gatcgagttt caggtgcggg ccattctggc tgtccccatc ctgcaaggca agaagctgtg
     1501 gggcctgctg gtggcccacc aactggccgg ccctcgggag tggcagacct gggaaatcga
     1561 cttcctgaaa cagcaagccg tggtgatggg catcgccatc cagcagagct aggaattcga
     1621 tatcaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     1681 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     1741 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     1801 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     1861 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     1921 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     1981 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
     2041 ctttccttgg ctgctcgcct atgttgccac ctggattctg cgcgggacgt ccttctgcta
     2101 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     2161 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     2221 cccgcatcga taccgagcgc tgctcgagag atctacgggt ggcatccctg tgacccctcc
     2281 ccagtgcctc tcctggccct ggaagttgcc actccagtgc ccaccagcct tgtcctaata
     2341 aaattaagtt gcatcatttt gtctgactag gtgtccttct ataatattat ggggtggagg
     2401 ggggtggtat ggagcaaggg gcaagttggg aagacaacct gtagggcctg cggggtctat
     2461 tgggaaccaa gctggagtgc agtggcacaa tcttggctca ctgcaatctc cgcctcctgg
     2521 gttcaagcga ttctcctgcc tcagcctccc gagttgttgg gattccaggc atgcatgacc
     2581 aggctcagct aatttttgtt tttttggtag agacggggtt tcaccatatt ggccaggctg
     2641 gtctccaact cctaatctca ggtgatctac ccaccttggc ctcccaaatt gctgggatta
     2701 caggcgtgaa ccactgctcc cttccctgtc cttctgattt tgtaggtaac cacgtgcgga
     2761 ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     2821 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     2881 cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta
     2941 cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag
     3001 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3061 cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3121 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     3181 cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata
     3241 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3301 aactggaaca acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc
     3361 gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     3421 caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc
     3481 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     3541 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     3601 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     3661 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     3721 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     3781 gagacaataa ccctgataaa tgcttcaata atatttaaaa aggaagagta tgagtattca
     3841 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     3901 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     3961 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     4021 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     4081 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     4141 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     4201 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     4261 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     4321 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     4381 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     4441 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     4501 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     4561 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     4621 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     4681 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     4741 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     4801 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     4861 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     4921 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     4981 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     5041 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     5101 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     5161 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     5221 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     5281 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     5341 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     5401 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     5461 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
