LOCUS       Exported                6985 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 6985)
  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..6985
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        279..556
                     /label=chicken beta-actin promoter
     primer_bind     954..1002
                     /label=0901-TD-F
     misc_feature    977..982
                     /label=Kozak sequence
     CDS             983..2422
                     /codon_start=1
                     /label=td8ox2StayGold
                     /translation="MASTGEELFTGVVPFKFQLKGTINGKSFTVEGEGEGNSHEGSHKG
                     KYVCTSGKLPMSWAALGTSFGYGMKYYTKYPSGLKNWFHEVMPEGFTYDRHIQYKGDGS
                     IHAKHQHFMKNGTYHNIVEFTGQDFKENSPVLTGDMNVSLPNEVQHIPRDDGVECPVTL
                     LYPLLSDKSKCVEAFQNTIIKPLHNQPAPDVPYHWIRKQYTQSKDDTEERDHIIQSETL
                     EAHLYSRTKLEGHGTGSTGSGSSGTASSEDNNMASTGEELFTGVVPFKFQLKGTINGKS
                     FTVEGEGEGNSHEGSHKGKYVCTSGKLPMSWAALGTSFGYGMKYYTKYPSGLKNWFHEV
                     MPEGFTYDRHIQYKGDGSIHAKHQHFMKNGTYHNIVEFTGQDFKENSPVLTGDMNVSLP
                     NEVQHIPRDDGVECPVTLLYPLLSDKSKCVEAFQNTIIKPLHNQPAPDVPYHWIRKQYT
                     QSKDDTEERDHIIQSETLEAHL"
     misc_feature    983..1660
                     /label=oxStayGold
     misc_feature    995..1021
                     /label=n1
     primer_bind     1657..1682
                     /label=TD-F
     misc_feature    1661..1681
                     /label=PT
     misc_feature    1682..1744
                     /label=Tdtomato
     primer_bind     complement(1731..1752)
                     /label=TD-R
     misc_feature    1745..2422
                     /label=oxStayGold
     CDS             2423..2452
                     /codon_start=1
                     /label=c4
                     /translation="PWHEPSASAV"
     primer_bind     complement(2433..2485)
                     /label=0901-TD-R
     CDS             2477..2533
                     /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             2546..3241
                     /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(3221..3265)
                     /label=0714-FusionRed-R
     misc_feature    3269..3857
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(3740..3751)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3900..4021
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   4183..4312
                     /label=AAV2 ITR
     rep_origin      complement(4549..5137)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5308..6168)
                     /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(6169..6273)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6299..6754)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     repeat_region   6826..6955
                     /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 ccatggctag cactggggaa gaactgttca caggagtcgt
     1021 gccgtttaag ttccagttga agggcacgat caatggaaag tctttcacag tggaaggaga
     1081 gggtgagggt aatagccatg aaggttcaca caaaggcaag tacgtctgta cctctggaaa
     1141 gcttccgatg tcatgggccg ctctgggtac ctcctttggg tatggcatga aatactatac
     1201 aaagtacccc tcagggttga agaactggtt ccatgaggtc atgcccgaag gattcaccta
     1261 tgatcgccac atccagtata agggcgacgg ctctattcat gccaaacatc aacacttcat
     1321 gaagaacggc acctaccata acatcgtgga gtttactgga caagacttta aggagaatag
     1381 cccagttctg accggtgata tgaacgtaag cctgcctaac gaagtgcagc acattcccag
     1441 ggacgatggg gttgagtgtc ctgttaccct gctgtaccct ctgttgagcg acaaatccaa
     1501 atgcgtagag gcgtttcaga acaccattat caaaccactg cataaccaac cagctcccga
     1561 tgtaccgtac cattggatac ggaaacagta tacccagtca aaagacgaca ctgaggaacg
     1621 tgaccacatt atccagtccg agacacttga agcacacctc tattcacgaa caaaactgga
     1681 ggggcatggc actggctcta ctggatccgg ctccagtgga acagcatcaa gcgaggacaa
     1741 caatatggcc tccacaggag aggagctgtt caccggagtc gttccattca aatttcagct
     1801 gaaaggcacc attaatggga aaagcttcac agtggaggga gaaggtgaag gtaatagtca
     1861 cgaagggagc cacaagggaa aatacgtctg cactagcggg aaactcccaa tgtcttgggc
     1921 tgcactgggg actagctttg gctacgggat gaagtactac acgaagtatc cttccggcct
     1981 caagaactgg tttcacgagg tgatgccaga agggtttacc tatgatagac acatacagta
     2041 caagggcgac ggcagtatcc acgcgaaaca ccagcatttc atgaagaatg ggacgtacca
     2101 caacattgtg gaatttacgg gccaagactt caaggagaat tctcccgtgc ttaccggcga
     2161 tatgaacgtg agtctgccaa atgaggtgca gcatatacct agggacgatg gtgtggaatg
     2221 tcccgtcaca ctgctgtatc ctctgctcag tgataaatct aagtgcgttg aggccttcca
     2281 aaacaccatc atcaagccct tgcataatca gcctgcccct gatgtgccct atcactggat
     2341 ccggaagcag tacacacagt ccaaggatga cactgaggag agagatcaca ttatccagag
     2401 cgagactctc gaagcccacc ttccatggca cgagccttct gcttctgccg tgcgaccggt
     2461 tcacgtggga agtggggcta caaacttctc actgctgaaa caggccgggg atgtggagga
     2521 aaacccagga ccacacgtga ctagtatggt gagcgagctg attaaggaga acatgcccat
     2581 gaagctgtac atggagggca ccgtgaacaa ccaccacttc aagtgcacat ccgagggcga
     2641 aggcaagccc tacgagggca cccagaccat gagaatcaag gtcgtcgagg gcggccctct
     2701 ccccttcgcc ttcgacatcc tggctaccag cttcatgtac ggcagcagaa ccttcatcaa
     2761 gcaccctccg ggcatccccg acttctttaa gcagtccttc cctgagggct tcacatggga
     2821 gagagtcacc acatacgaag acgggggcgt gctgaccgct acccaggaca ccagcctcca
     2881 ggacggctgc ctcatctaca acgtcaaggt tagaggggtg aacttcccag ccaacggccc
     2941 tgtgatgcag aagaaaacac tcggctggga ggcctccacc gagacgatgt accctgctga
     3001 cggaggactg gaaggcgcat gtgacatggc tctgaagctc gtgggaggag gacacctgat
     3061 ctgcaacctt gagacaacat acagatccaa gaaacccgct acgaacctca agatgcccgg
     3121 cgtctacaac gtggaccaca gactggaaag aatcaaggag gccgacgatg agacatacgt
     3181 cgagcagcac gaggtggctg tggccagata ctctactggt ggcgctggtg atggaggtaa
     3241 ataagaattc gatatcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag
     3301 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     3361 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     3421 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     3481 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     3541 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     3601 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     3661 gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     3721 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     3781 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     3841 ttgggccgcc tccccgcatc gataccgtcg acccgggcgg ccgcttcgag cagacatgat
     3901 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     3961 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     4021 taacaacaac aattgcattc attttatgtt tcaggttcag ggggagatgt gggaggtttt
     4081 ttaaagcaag taaaacctct acaaatgtgg taaaatcgat aaggatcttc ctagagcatg
     4141 gctacgtaga taagtagcat ggcgggttaa tcattaacta caaggaaccc ctagtgatgg
     4201 agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg
     4261 cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcatta
     4321 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     4381 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     4441 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     4501 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     4561 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     4621 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     4681 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     4741 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     4801 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     4861 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     4921 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     4981 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     5041 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     5101 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     5161 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     5221 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     5281 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     5341 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     5401 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     5461 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     5521 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     5581 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     5641 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     5701 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     5761 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     5821 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     5881 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     5941 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     6001 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     6061 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     6121 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     6181 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     6241 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa
     6301 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     6361 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     6421 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     6481 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     6541 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     6601 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     6661 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     6721 cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt cgccattcag gctgcgcaac
     6781 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctgcg cgctcgctcg
     6841 ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg cccggcctca
     6901 gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg ttccttgtag
     6961 ttaatgatta acccgccatg ctact
//
