pAAV-CAG-iRFP670-P2A-GFP

LOCUS       pAAV-CAG-iRFP670-P2A-GFP               6466 bp DNA     circular SYN 10-OCT-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6466)
  AUTHORS   Triple Threat
  TITLE     Direct Submission
  JOURNAL   Exported Jun 22, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..6466
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    215..1155
                     /label=ColE1
                     /note="ColE1"
     rep_origin      complement(397..985)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1156..2016)
                     /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"
     promoter        complement(2017..2121)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2147..2602)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     polyA_signal    2965..3086
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    3129..3717
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3235..3246
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             complement(3742..4458)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     4369..4387
                     /label=EGFP-R
     CDS             complement(4471..4527)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             complement(4552..5484)
                     /codon_start=1
                     /product="near-infrared fluorescent protein with an 
                     emission peak at 670 nm, engineered from a bacterial 
                     phytochrome (Shcherbakova and Verkhusha, 2013)"
                     /label=iRFP670
                     /note="derived from Rhodopseudomonas palustris RpBphP6"
                     /translation="MARKVDLTSCDREPIHIPGSIQPCGCLLACDAQAVRITRITENAG
                     AFFGRETPRVGELLADYFGETEAHALRNALAQSSDPKRPALIFGWRDGLTGRTFDISLH
                     RHDGTSIIEFEPAAAEQADNPLRLTRQIIARTKELKSLEEMAARVPRYLQAMLGYHRVM
                     LYRFADDGSGMVIGEAKRSDLESFLGQHFPASLVPQQARLLYLKNAIRVVSDSRGISSR
                     IVPEHDASGAALDLSFAHLRSISPCHLEFLRNMGVSASMSLSIIIDGTLWGLIICHHYE
                     PRAVPMAQRVAAEMFADFLSLHFTAAHHQR"
     regulatory      5485..5490
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5911..6188)
                     /label=chicken beta-actin promoter
ORIGIN
        1 agtagcatgg cgggttaatc attaactaca aggaacccct agtgatggag ttggccactc
       61 cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg
      121 gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcattaat gaatcggcca
      181 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
      241 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
      301 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
      361 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
      421 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
      481 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
      541 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
      601 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
      661 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
      721 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
      781 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
      841 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
      901 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
      961 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     1021 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     1081 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     1141 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     1201 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     1261 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     1321 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     1381 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     1441 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     1501 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     1561 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     1621 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     1681 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     1741 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     1801 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     1861 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     1921 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     1981 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     2041 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     2101 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt
     2161 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     2221 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     2281 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     2341 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     2401 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     2461 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     2521 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     2581 aatgcgccgc tacagggcgc gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg
     2641 cgatcggtgc gggcctcttc gctattacgc cagctgcgcg ctcgctcgct cactgaggcc
     2701 gcccgggcaa agcccgggcg tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga
     2761 gcgcgcagag agggagtggc caactccatc actaggggtt ccttgtagtt aatgattaac
     2821 ccgccatgct acttatctac gtagccatgc tctaggaaga tccttatcga ttttaccaca
     2881 tttgtagagg ttttacttgc tttaaaaaac ctcccacatc tccccctgaa cctgaaacat
     2941 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
     3001 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     3061 ttgtccaaac tcatcaatgt atcttatcat gtctgctcga agcggccgcc cgggtcgacg
     3121 gtatcgatgc ggggaggcgg cccaaaggga gatccgactc gtctgagggc gaaggcgaag
     3181 acgcggaaga ggccgcagag ccggcagcag gccgcgggaa ggaaggtccg ctggattgag
     3241 ggccgaaggg acgtagcaga aggacgtccc gcgcagaatc caggtggcaa cacaggcgag
     3301 cagccaagga aaggacgatg atttccccga caacaccacg gaattgtcag tgcccaacag
     3361 ccgagcccct gtccagcagc gggcaaggca ggcggcgatg agttccgccg tggcaatagg
     3421 gagggggaaa gcgaaagtcc cggaaaggag ctgacaggtg gtggcaatgc cccaaccagt
     3481 gggggttgcg tcagcaaaca cagtgcacac cacgccacgt tgcctgacaa cgggccacaa
     3541 ctcctcataa agagacagca accaggattt atacaaggag gagaaaatga aagccatacg
     3601 ggaagcaata gcatgataca aaggcattaa agcagcgtat ccacatagcg taaaaggagc
     3661 aacatagtta agaataccag tcaatctttc acaaattttg taatccagag gttgattatc
     3721 gataagcttg atatcgaatt cttacttgta cagctcgtcc atgccgagag tgatcccggc
     3781 ggcggtcacg aactccagca ggaccatgtg atcgcgcttc tcgttggggt ctttgctcag
     3841 ggcggactgg gtgctcaggt agtggttgtc gggcagcagc acggggccgt cgccgatggg
     3901 ggtgttctgc tggtagtggt cggcgagctg cacgctgccg tcctcgatgt tgtggcggat
     3961 cttgaagttc accttgatgc cgttcttctg cttgtcggcc atgatataga cgttgtggct
     4021 gttgtagttg tactccagct tgtgccccag gatgttgccg tcctccttga agtcgatgcc
     4081 cttcagctcg atgcggttca ccagggtgtc gccctcgaac ttcacctcgg cgcgggtctt
     4141 gtagttgccg tcgtccttga agaagatggt gcgctcctgg acgtagcctt cgggcatggc
     4201 ggacttgaag aagtcgtgct gcttcatgtg gtcggggtag cggctgaagc actgcacgcc
     4261 gtaggtcagg gtggtcacga gggtgggcca gggcacgggc agcttgccgg tggtgcagat
     4321 gaacttcagg gtcagcttgc cgtaggtggc atcgccctcg ccctcgccgg acacgctgaa
     4381 cttgtggccg tttacgtcgc cgtccagctc gaccaggatg ggcaccaccc cggtgaacag
     4441 ctcctcgccc ttgctcacac tagtcacgtg tggtcctggg ttttcctcca catccccggc
     4501 ctgtttcagc agtgagaagt ttgtagcccc acttcccacg tgaaccggtc ggcgttggtg
     4561 gtgggcggcg gtgaagtgca gcgataagaa gtcggcgaac atttcggccg cgacgcgctg
     4621 cgccatcggc acggcacgcg gctcgtaatg atgacagatg atcaatcccc atagcgtgcc
     4681 gtcaatgatg atcgacagcg acatcgaggc gctgacgccc atgttccgca gaaattcgag
     4741 atggcagggc gagatgctgc gcaggtgcgc gaacgacaga tcgagcgcgg cgccggaggc
     4801 gtcgtgctcg ggcacgatcc ggctgctgat gccgcgcgaa tccgagacca cgcggatcgc
     4861 gttcttcaag tacagtagcc gcgcctgctg cgggaccagc gacgccggaa agtgctgacc
     4921 gagaaagctc tcgaggtcgc tgcgcttcgc ctcgccgatc accatcccgg agccgtcgtc
     4981 cgcgaagcgg tacaacatca cgcggtgata gccgagcatc gcctgcagat agcgcggcac
     5041 ccgtgcggcc atctcttcga gcgacttcag ttctttggtg cgcgcgatga tctgccgcgt
     5101 cagccgcagc ggattgtcgg cctgttcggc cgccgcaggc tcgaactcga tgatcgatgt
     5161 accgtcatgg cgatgcagtg agatgtcgaa ggtgcggccg gtcaggccgt cgcgccaacc
     5221 gaagatcagc gccggtcgct ttggatcgga ggactgcgcc agtgcgttgc gcagcgcatg
     5281 ggcttcggtc tcgccgaagt aatcggcgag tagctcaccg acccgcggag tttcgcgtcc
     5341 aaagaacgcg ccggcatttt ccgtaatgcg cgtgatccgc accgcctgcg cgtcgcaggc
     5401 tagcaggcag ccgcacggct gaatgctgcc ggggatgtgg atcggctcgc gatcgcagga
     5461 ggtgagatcg accttacgcg ccatggtggc ggtaccggat ccaattcttt gccaaaatga
     5521 tgagacagca caataaccag cacgttgccc aggagctgta ggaaaaagaa gaaggcatga
     5581 acatggttag cagaggctct agagccgccg gtcacacgcc agaagccgaa ccccgccctg
     5641 ccccgtcccc cccgaaggca gccgtccccc cgcggacagc cccgagccgc tccccccgca
     5701 caaagggccc tcccggagcc cctcaaggct ttcacgcagc cacagaaaag aaacaagccg
     5761 tcattaaacc aagcgctaat tacagcccgg aggagaaggg ccgtcccgcc cgctcacctg
     5821 tgggagtaac gcggtcagtc agagccgggg cgggcggcgc gaggcggcgg cggagcgggg
     5881 cacggggcga aggcagcgcg cagcgactcc cgcccgccgc gcgcttcgct ttttataggg
     5941 ccgccgccgc cgccgcctcg ccataaaagg aaactttcgg agcgcgccgc tctgattggc
     6001 tgccgccgca cctctccgcc tcgccccgcc ccgcccctcg ccccgccccg ccccgcctgg
     6061 cgcgcgcccc cccccccccc ccgcccccat cgctgcacaa aataattaaa aaataaataa
     6121 atacaaaatt gggggtgggg agggggggga gatggggaga gtgaagcaga acgtggggct
     6181 cacctcgacc atggtaatag cgatgactaa tacgtagatg tactgccaag taggaaagtc
     6241 ccataaggtc atgtactggg cataatgcca ggcgggccat ttaccgtcat tgacgtcaat
     6301 agggggcgta cttggcatat gatacacttg atgtactgcc aagtgggcag tttaccgtaa
     6361 atactccacc cattgacgtc aatggaaagt ccctattggc gttactatgg gaacatacgt
     6421 cattattgac gtcaatggta ctcttcctag agcatggcta cgtaga
//