LOCUS       Exported                5656 bp ds-DNA     circular SYN 14-MAY-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5656)
  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..5656
                     /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..1941)
                     /label=0503-emiRFP703-R
     misc_feature    1940..2528
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(2411..2422)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2571..2692
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(3220..3808)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3979..4839)
                     /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(4840..4944)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4970..5425)
                     /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 gctaagaatt
     1921 cgatatcaag cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg
     1981 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     2041 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     2101 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     2161 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     2221 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     2281 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc
     2341 gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     2401 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     2461 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     2521 ctccccgcat cgataccgtc gacccgggcg gccgcttcga gcagacatga taagatacat
     2581 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     2641 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     2701 caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt tttaaagcaa
     2761 gtaaaacctc tacaaatgtg gtaaaatcga taaggatctt cctagagcat ggctacgtag
     2821 ataagtagca tggcgggtta atcattaact acaaggaacc cctagtgatg gagttggcca
     2881 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc
     2941 cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcatt aatgaatcgg
     3001 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     3061 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     3121 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     3181 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     3241 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     3301 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     3361 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     3421 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     3481 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3541 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     3601 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     3661 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     3721 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     3781 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     3841 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     3901 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     3961 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     4021 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     4081 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     4141 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     4201 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     4261 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     4321 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     4381 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     4441 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     4501 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     4561 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     4621 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     4681 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     4741 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     4801 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     4861 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     4921 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc
     4981 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     5041 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     5101 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     5161 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     5221 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     5281 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     5341 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     5401 cttaatgcgc cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa
     5461 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgc gcgctcgctc gctcactgag
     5521 gccgcccggg caaagcccgg gcgtcgggcg acctttggtc gcccggcctc agtgagcgag
     5581 cgagcgcgca gagagggagt ggccaactcc atcactaggg gttccttgta gttaatgatt
     5641 aacccgccat gctact
//
