LOCUS       Exported                6211 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 6211)
  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..6211
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        279..556
                     /label=chicken beta-actin promoter
     primer_bind     951..1007
                     /label=0901-AAUSFP-F
     misc_feature    977..982
                     /label=Kozak sequence
     CDS             983..1678
                     /codon_start=1
                     /label=AausFP1
                     /translation="MSYGALLFREKIPYVVEMEGDVEGMKFSVRGKGHGDANTGKIEAS
                     FICTTGELPVPWSSILTTVTYGAQCFAKYPNDIKDYPKSAMPEGYVQERTITFENDGVY
                     KTRAEVTYEKGSVYNRVTLNGSGFKKGGNILGKKLEFNYNPHCIYVLPDVQNNGIKCYI
                     NIVHDVIGGGQIIAAHQQLNTPLGGGPVDIPHYHHIQAHTILSKDPKETRDHMNVVEVF
                     RAIDCKTAYA"
     primer_bind     complement(1657..1711)
                     /label=0901-AAUSFP-R
     CDS             1703..1759
                     /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             1772..2467
                     /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(2447..2491)
                     /label=0714-FusionRed-R
     misc_feature    2495..3083
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(2966..2977)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3126..3247
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   3409..3538
                     /label=AAV2 ITR
     rep_origin      complement(3775..4363)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4534..5394)
                     /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(5395..5499)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5525..5980)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     repeat_region   6052..6181
                     /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 ggtaccgcca ccatgagtta cggagcactt ttgttcagag aaaaaatccc
     1021 ctatgttgtt gagatggaag gtgatgttga gggaatgaag ttttcagtaa gaggtaaagg
     1081 acatggtgat gctaacactg gtaaaattga agccagtttc atctgcacaa ctggtgaact
     1141 accagtacct tggtcaagca ttctaacaac tgtcacatac ggtgcacagt gttttgcaaa
     1201 atatccaaat gatatcaaag actacccaaa aagtgctatg ccagaaggtt acgttcaaga
     1261 gcgaacaatc acttttgaaa acgacggcgt ttataagaca cgcgctgaag tcacgtacga
     1321 gaaaggatca gtttataacc gagttacgtt gaacggttca ggatttaaaa aaggcggtaa
     1381 catcttgggc aaaaaattgg aatttaacta caaccctcat tgcatctatg ttcttccaga
     1441 tgtacaaaat aatggaatca aatgctacat caacattgtt catgacgtaa ttggaggagg
     1501 acaaataatt gctgcgcacc aacaattaaa tactcctctt ggcggtggac cagtcgacat
     1561 tccacactac catcatatac aagcgcacac aatactttct aaagatccca aagaaacaag
     1621 agatcatatg aacgtagtag aggtgttcag agctatcgat tgcaaaaccg catatgcacg
     1681 accggttcac gtgggaagtg gggctacaaa cttctcactg ctgaaacagg ccggggatgt
     1741 ggaggaaaac ccaggaccac acgtgactag tatggtgagc gagctgatta aggagaacat
     1801 gcccatgaag ctgtacatgg agggcaccgt gaacaaccac cacttcaagt gcacatccga
     1861 gggcgaaggc aagccctacg agggcaccca gaccatgaga atcaaggtcg tcgagggcgg
     1921 ccctctcccc ttcgccttcg acatcctggc taccagcttc atgtacggca gcagaacctt
     1981 catcaagcac cctccgggca tccccgactt ctttaagcag tccttccctg agggcttcac
     2041 atgggagaga gtcaccacat acgaagacgg gggcgtgctg accgctaccc aggacaccag
     2101 cctccaggac ggctgcctca tctacaacgt caaggttaga ggggtgaact tcccagccaa
     2161 cggccctgtg atgcagaaga aaacactcgg ctgggaggcc tccaccgaga cgatgtaccc
     2221 tgctgacgga ggactggaag gcgcatgtga catggctctg aagctcgtgg gaggaggaca
     2281 cctgatctgc aaccttgaga caacatacag atccaagaaa cccgctacga acctcaagat
     2341 gcccggcgtc tacaacgtgg accacagact ggaaagaatc aaggaggccg acgatgagac
     2401 atacgtcgag cagcacgagg tggctgtggc cagatactct actggtggcg ctggtgatgg
     2461 aggtaaataa gaattcgata tcaagcttat cgataatcaa cctctggatt acaaaatttg
     2521 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     2581 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     2641 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     2701 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     2761 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     2821 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     2881 gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg
     2941 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     3001 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     3061 ctccctttgg gccgcctccc cgcatcgata ccgtcgaccc gggcggccgc ttcgagcaga
     3121 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     3181 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     3241 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga
     3301 ggttttttaa agcaagtaaa acctctacaa atgtggtaaa atcgataagg atcttcctag
     3361 agcatggcta cgtagataag tagcatggcg ggttaatcat taactacaag gaacccctag
     3421 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     3481 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct
     3541 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     3601 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     3661 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     3721 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     3781 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     3841 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     3901 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     3961 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     4021 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     4081 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     4141 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     4201 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     4261 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     4321 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     4381 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     4441 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     4501 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     4561 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     4621 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     4681 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     4741 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     4801 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     4861 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     4921 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     4981 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     5041 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     5101 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     5161 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     5221 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     5281 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     5341 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     5401 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     5461 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     5521 acctaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     5581 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     5641 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     5701 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     5761 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     5821 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     5881 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     5941 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc attcaggctg
     6001 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctgcgcgct
     6061 cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc gggcgacctt tggtcgcccg
     6121 gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc
     6181 ttgtagttaa tgattaaccc gccatgctac t
//
