LOCUS       Exported                6466 bp ds-DNA     circular SYN 04-MAR-2022
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 Friday, Jan 20, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6466
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        279..556
                     /label=chicken beta-actin promoter
     primer_bind     962..1004
                     /label=0117-iRFP670-F
     regulatory      977..986
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             983..1915
                     /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=miRFP670-2
                     /note="derived from Rhodopseudomonas palustris RpBphP6"
                     /translation="MARKVDLTSCDREPIHIPGSIQPCGCLLACDAQAVRITRITENAG
                     AFFGRETPRVGELLADYFGETEAHALRNALAQSSDPKRPALIFGWRDGLTGRTFDISLH
                     RHDGTSIIEFEPAAAEQADNPLRLTRQIIARTKELKSLEEMAARVPRYLQAMLGYHRVM
                     LYRFADDGSGMVIGEAKRSDLESFLGQHFPASLVPQQARLLYLKNAIRVVSDSRGISSR
                     IVPEHDASGAALDLSFAHLRSISPCHLEFLRNMGVSASMSLSIIIDGTLWGLIICHHYE
                     PRAVPMAQRVAAKRFAERLSTHFTAAHHQR"
     CDS             1940..1996
                     /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             2009..2725
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     misc_feature    2750..3338
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(3221..3232)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3381..3502
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(4030..4618)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4789..5649)
                     /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(5650..5754)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5780..6235)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 tctacgtagc catgctctag gaagagtacc attgacgtca ataatgacgt atgttcccat
       61 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      121 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
      181 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
      241 gcagtacatc tacgtattag tcatcgctat taccatggtc gaggtgagcc ccacgttctg
      301 cttcactctc cccatctccc ccccctcccc acccccaatt ttgtatttat ttatttttta
      361 attattttgt gcagcgatgg gggcgggggg gggggggggg cgcgcgccag gcggggcggg
      421 gcggggcgag gggcggggcg gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg
      481 cgcgctccga aagtttcctt ttatggcgag gcggcggcgg cggcggccct ataaaaagcg
      541 aagcgcgcgg cgggcgggag tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc
      601 cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta ctcccacagg tgagcgggcg
      661 ggacggccct tctcctccgg gctgtaatta gcgcttggtt taatgacggc ttgtttcttt
      721 tctgtggctg cgtgaaagcc ttgaggggct ccgggagggc cctttgtgcg gggggagcgg
      781 ctcggggctg tccgcggggg gacggctgcc ttcggggggg acggggcagg gcggggttcg
      841 gcttctggcg tgtgaccggc ggctctagag cctctgctaa ccatgttcat gccttcttct
      901 ttttcctaca gctcctgggc aacgtgctgg ttattgtgct gtctcatcat tttggcaaag
      961 aattggatcc ggtaccgcca ccatggcgcg taaggtcgat ctcacctcct gcgatcgcga
     1021 gccgatccac atccccggca gcattcagcc gtgcggctgc ctgctagcct gcgacgcgca
     1081 ggcggtgcgg atcacgcgca ttacggaaaa tgccggcgcg ttctttggac gcgaaactcc
     1141 gcgggtcggt gagctactcg ccgattactt cggcgagacc gaagcccatg cgctgcgcaa
     1201 cgcactggcg cagtcctccg atccaaagcg accggcgctg atcttcggtt ggcgcgacgg
     1261 cctgaccggc cgcaccttcg acatctcact gcatcgccat gacggtacat cgatcatcga
     1321 gttcgagcct gcggcggccg aacaggccga caatccgctg cggctgacgc ggcagatcat
     1381 cgcgcgcacc aaagaactga agtcgctcga agagatggcc gcacgggtgc cgcgctatct
     1441 gcaggcgatg ctcggctatc accgcgtgat gttgtaccgc ttcgcggacg acggctccgg
     1501 gatggtgatc ggcgaggcga agcgcagcga cctcgagagc tttctcggtc agcactttcc
     1561 ggcgtcgctg gtcccgcagc aggcgcggct actgtacttg aagaacgcga tccgcgtggt
     1621 ctcggattcg cgcggcatca gcagccggat cgtgcccgag cacgacgcct ccggcgccgc
     1681 gctcgatctg tcgttcgcgc acctgcgcag catctcgccc tgccatctcg aatttctgcg
     1741 gaacatgggc gtcagcgcct cgatgtcgct gtcgatcatc attgacggca cgctatgggg
     1801 attgatcatc tgtcatcatt acgagccgcg tgccgtgccg atggcgcagc gcgtcgcggc
     1861 caaaaggttc gccgagcgct tatcgacgca cttcaccgcc gcccaccacc aacgccgacc
     1921 ggttcacgtg ggaagtgggg ctacaaactt ctcactgctg aaacaggccg gggatgtgga
     1981 ggaaaaccca ggaccacacg tgactagtgt gagcaagggc gaggagctgt tcaccggggt
     2041 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
     2101 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
     2161 caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt
     2221 cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     2281 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
     2341 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     2401 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta
     2461 tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat
     2521 cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     2581 ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc
     2641 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
     2701 cggcatggac gagctgtaca agtaagaatt cgatatcaag cttatcgata atcaacctct
     2761 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     2821 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     2881 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     2941 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     3001 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     3061 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     3121 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc
     3181 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     3241 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     3301 tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgtc gacccgggcg
     3361 gccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     3421 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     3481 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     3541 gggggagatg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga
     3601 taaggatctt cctagagcat ggctacgtag ataagtagca tggcgggtta atcattaact
     3661 acaaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     3721 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg
     3781 agcgagcgcg cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     3841 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     3901 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     3961 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     4021 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     4081 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     4141 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     4201 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     4261 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     4321 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     4381 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     4441 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     4501 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     4561 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     4621 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     4681 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     4741 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     4801 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     4861 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     4921 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     4981 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     5041 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     5101 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     5161 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     5221 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     5281 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     5341 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     5401 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     5461 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     5521 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     5581 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     5641 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     5701 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     5761 tccccgaaaa gtgccaccta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     5821 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     5881 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     5941 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     6001 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     6061 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     6121 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     6181 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat
     6241 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     6301 cgccagctgc gcgctcgctc gctcactgag gccgcccggg caaagcccgg gcgtcgggcg
     6361 acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc
     6421 atcactaggg gttccttgta gttaatgatt aacccgccat gctact
//
