LOCUS       Exported                6236 bp ds-DNA     circular SYN 01-NOV-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6236)
  AUTHORS   Triple Threat
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Nov 6, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6236
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        279..556
                     /label=chicken beta-actin promoter
     primer_bind     951..1006
                     /label=0907-1040-F
     misc_feature    981..986
                     /label=Kozak sequence
     CDS             987..1703
                     /codon_start=1
                     /product="bright green-yellow fluorescent protein derived 
                     from mClover2, with improved photostability (Bajar et al., 
                     2003)"
                     /label=mClover3
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHYVYITADKQKNCIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(1683..1731)
                     /label=0907-1040-R
     CDS             1728..1784
                     /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             1797..2492
                     /codon_start=1
                     /product="monomeric red fluorescent protein optimized for 
                     use as a fusion tag (Shemiakina et al., 2012)"
                     /label=FusionRed
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMPMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     VVEGGPLPFAFDILATSFMYGSRTFIKHPPGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKVRGVNFPANGPVMQKKTLGWEASTETMYPADGGLEGACDMAL
                     KLVGGGHLICNLETTYRSKKPATNLKMPGVYNVDHRLERIKEADDETYVEQHEVAVARY
                     STGGAGDGGK"
     primer_bind     complement(2472..2516)
                     /label=0714-FusionRed-R
     misc_feature    2520..3108
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(2991..3002)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3151..3272
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   3434..3563
                     /label=AAV2 ITR
     rep_origin      complement(3800..4388)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4559..5419)
                     /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(5420..5524)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5550..6005)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     repeat_region   6077..6206
                     /label=AAV2 ITR
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 ggtaccataa gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg
     1021 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtccgcg
     1081 gcgagggcga gggcgatgcc accaacggca agctgaccct gaagttcatc tgcaccaccg
     1141 gcaagctgcc cgtgccctgg cccaccctcg tgaccacctt cggctacggc gtggcctgct
     1201 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     1261 gctacgtcca ggagcgcacc atctctttca aggacgacgg tacctacaag acccgcgccg
     1321 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     1381 aggaggacgg caacatcctg gggcacaagc tggagtacaa cttcaacagc cactacgtct
     1441 atatcacggc cgacaagcag aagaactgca tcaaggctaa cttcaagatc cgccacaacg
     1501 ttgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     1561 gccccgtgct gctgcccgac aaccactacc tgagccatca gtccaagctg agcaaagacc
     1621 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggattacac
     1681 atggcatgga cgagctgtac aagcgaccgg ttcacgtggg aagtggggct acaaacttct
     1741 cactgctgaa acaggccggg gatgtggagg aaaacccagg accacacgtg actagtatgg
     1801 tgagcgagct gattaaggag aacatgccca tgaagctgta catggagggc accgtgaaca
     1861 accaccactt caagtgcaca tccgagggcg aaggcaagcc ctacgagggc acccagacca
     1921 tgagaatcaa ggtcgtcgag ggcggccctc tccccttcgc cttcgacatc ctggctacca
     1981 gcttcatgta cggcagcaga accttcatca agcaccctcc gggcatcccc gacttcttta
     2041 agcagtcctt ccctgagggc ttcacatggg agagagtcac cacatacgaa gacgggggcg
     2101 tgctgaccgc tacccaggac accagcctcc aggacggctg cctcatctac aacgtcaagg
     2161 ttagaggggt gaacttccca gccaacggcc ctgtgatgca gaagaaaaca ctcggctggg
     2221 aggcctccac cgagacgatg taccctgctg acggaggact ggaaggcgca tgtgacatgg
     2281 ctctgaagct cgtgggagga ggacacctga tctgcaacct tgagacaaca tacagatcca
     2341 agaaacccgc tacgaacctc aagatgcccg gcgtctacaa cgtggaccac agactggaaa
     2401 gaatcaagga ggccgacgat gagacatacg tcgagcagca cgaggtggct gtggccagat
     2461 actctactgg tggcgctggt gatggaggta aataagaatt cgatatcaag cttatcgata
     2521 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     2581 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     2641 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     2701 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     2761 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     2821 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     2881 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     2941 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     3001 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     3061 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgtc
     3121 gacccgggcg gccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca
     3181 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     3241 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     3301 ttcaggttca gggggagatg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg
     3361 gtaaaatcga taaggatctt cctagagcat ggctacgtag ataagtagca tggcgggtta
     3421 atcattaact acaaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     3481 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
     3541 tcagtgagcg agcgagcgcg cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     3601 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     3661 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     3721 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     3781 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     3841 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     3901 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     3961 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     4021 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     4081 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     4141 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     4201 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     4261 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     4321 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     4381 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     4441 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     4501 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     4561 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     4621 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     4681 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     4741 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     4801 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     4861 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     4921 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     4981 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     5041 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     5101 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     5161 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     5221 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     5281 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     5341 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     5401 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     5461 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     5521 cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc gttaatattt tgttaaaatt
     5581 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     5641 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     5701 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     5761 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     5821 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     5881 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     5941 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     6001 cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     6061 ttcgctatta cgccagctgc gcgctcgctc gctcactgag gccgcccggg caaagcccgg
     6121 gcgtcgggcg acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt
     6181 ggccaactcc atcactaggg gttccttgta gttaatgatt aacccgccat gctact
//
