LOCUS       Exported                6463 bp ds-DNA     circular SYN 19-DEC-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6463)
  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..6463
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        279..556
                     /label=chicken beta-actin promoter
     primer_bind     956..1009
                     /label=PM349-F-second
     regulatory      977..986
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             983..1912
                     /codon_start=1
                     /product="bright monomeric near-infrared fluorescent 
                     protein engineered from a bacterial phytochrome, with an 
                     emission maximum of 703 nm (Shcherbakova et al., 2016)"
                     /label=emiRFP703
                     /label=ORF frame 1
                     /translation="MAEGSVARQPDLLTCEHEEIHLAGSIQPHGALLVVSEHDHRVIQA
                     SANAAEFLNLGSVLGVPLAEIDGDLLIKILPHLDPTAEGMPVAVRCRIGNPSTEYCGLM
                     HRPPEGGLIIELERAGPSIDLSGTLAPALERIRTAGSLRALCDDTVLLFQQCTGYDRVM
                     VYRFDEQGHGLVFSECHVPGLESYFGNRYPSSLVPQMARQLYVRQRVRVLVDVTYQPVP
                     LEPRLSPLTGRDLDMSGCFLRSMSPIHLQFLKDMGVRATLAVSLVVGGKLWGLVVCHHY
                     LPRFIRFELRAICKRLAERIATRITALES"
     CDS             983..1024
                     /codon_start=1
                     /label=emiRFP2
                     /translation="MAEGSVARQPDLLT"
     primer_bind     990..1048
                     /label=PM349-F-first
     primer_bind     complement(1893..1935)
                     /label=0120-emiRFP703-R
     CDS             1937..1993
                     /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             2006..2722
                     /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     complement(2699..2743)
                     /label=PM349-R
     misc_feature    2747..3335
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(3218..3229)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3378..3499
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(4027..4615)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4786..5646)
                     /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(5647..5751)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5777..6232)
                     /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 ccatggccga gggcagcgtg gccagacagc ccgacctgct
     1021 gacctgcgaa catgaagaga tccacctcgc cggctcgatc cagccgcatg gcgcgcttct
     1081 ggtcgtcagc gaacatgatc atcgcgtcat ccaggccagc gccaacgccg cggaatttct
     1141 gaatctcgga agcgtactcg gcgttccgct cgccgagatc gacggcgatc tgttgatcaa
     1201 gatcctaccg catctcgatc ccaccgccga aggcatgccg gtcgcggtgc gctgccggat
     1261 cggcaatccc tctacggagt actgcggtct gatgcatcgg cctccggaag gcgggctgat
     1321 catcgaactc gaacgtgccg gcccgtcgat cgatctgtca ggcacgctgg cgccggcgct
     1381 ggagcggatc cgcacggcgg gttcactgcg cgcgctgtgc gatgacaccg tgctgctgtt
     1441 tcagcagtgc accggctacg accgggtgat ggtgtatcgt ttcgatgagc aaggccacgg
     1501 cctggtattc tccgagtgcc atgtgcctgg gctcgaatcc tatttcggca accgctatcc
     1561 gtcgtcgctg gtcccgcaga tggcgcggca gctgtacgtg cggcagcgcg tccgcgtgct
     1621 ggtcgacgtc acctatcagc cggtgccgct ggagccgcgg ctgtcgccgc tgaccgggcg
     1681 cgatctcgac atgtcgggct gcttcctgcg ctcgatgtcg ccgatccatc tgcagttcct
     1741 gaaggacatg ggcgtgcgcg ccaccctggc ggtgtcgctg gtggtcggcg gcaagctgtg
     1801 gggcctggtt gtctgtcacc attatctgcc gcgcttcatc cgtttcgagc tgcgggcgat
     1861 ctgcaaacgg ctcgccgaaa ggatcgcgac gcggatcacc gcgcttgaga gccgaccggt
     1921 tcacgtggga agtggggcta caaacttctc actgctgaaa caggccgggg atgtggagga
     1981 aaacccagga ccacacgtga ctagtgtgag caagggcgag gagctgttca ccggggtggt
     2041 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     2101 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     2161 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
     2221 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     2281 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     2341 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     2401 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     2461 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
     2521 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     2581 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
     2641 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     2701 catggacgag ctgtacaagt aagaattcga tatcaagctt atcgataatc aacctctgga
     2761 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     2821 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     2881 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     2941 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     3001 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     3061 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     3121 ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac
     3181 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     3241 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     3301 gacgagtcgg atctcccttt gggccgcctc cccgcatcga taccgtcgac ccgggcggcc
     3361 gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     3421 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     3481 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     3541 ggagatgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta aaatcgataa
     3601 ggatcttcct agagcatggc tacgtagata agtagcatgg cgggttaatc attaactaca
     3661 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
     3721 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc
     3781 gagcgcgcag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     3841 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     3901 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     3961 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     4021 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4081 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4141 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4201 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4261 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4321 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4381 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4441 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     4501 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     4561 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     4621 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     4681 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     4741 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     4801 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     4861 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     4921 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     4981 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     5041 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     5101 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     5161 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     5221 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     5281 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     5341 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     5401 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     5461 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     5521 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     5581 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     5641 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     5701 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     5761 ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     5821 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     5881 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     5941 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     6001 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     6061 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     6121 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     6181 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg
     6241 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     6301 cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc
     6361 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc
     6421 actaggggtt ccttgtagtt aatgattaac ccgccatgct act
//
