LOCUS       Exported                6217 bp ds-DNA     circular SYN 06-NOV-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6217)
  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..6217
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        279..556
                     /label=chicken beta-actin promoter
     primer_bind     956..1004
                     /label=0711-PM960-F
     misc_feature    977..982
                     /label=Kozak sequence
     CDS             983..1684
                     /codon_start=1
                     /label=mBaoJin
                     /translation="MVSKGEEENMASTPFKFQLKGTINGKSFTVEGEGEGNSHEGSHKG
                     KYVCTSGKLPMSWAALGTTFGYGMKYYTKYPSGLKNWFREVMPGGFTYDRHIQYKGDGS
                     IHAKHQHFMKNGTYHNIVEFTGQDFKENSPVLTGDMNVSLPNEVPQIPRDDGVECPVTL
                     LYPLLSDKSKYVEAHQYTICKPLHNQPAPDVPYHWIRKQYTQSKDDAEERDHICQSETL
                     EAHLKGMDELYK"
     primer_bind     1009..1043
                     /label=0714-StayBlue-F
     primer_bind     complement(1633..1657)
                     /label=0714-StayBlue-R
     primer_bind     complement(1663..1707)
                     /label=0711-PM960-R
     CDS             1709..1765
                     /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             1778..2473
                     /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(1954..1974)
                     /label=FusionRed-Rseq
     primer_bind     complement(2453..2497)
                     /label=0714-FusionRed-R
     misc_feature    2501..3089
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(2972..2983)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3132..3253
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   3415..3544
                     /label=AAV2 ITR
     rep_origin      complement(3781..4369)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4540..5400)
                     /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(5401..5505)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5531..5986)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     repeat_region   6058..6187
                     /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 ccatggtctc aaagggagag gaggaaaaca tggctagtac
     1021 accatttaaa tttcaactta aaggaaccat caatggcaaa tcgtttaccg ttgaaggcga
     1081 aggtgaaggg aactcacatg aaggttctca taaaggaaaa tatgtttgta caagtggaaa
     1141 actaccgatg tcatgggcag cacttgggac aacctttggt tatggaatga aatattatac
     1201 caaatatcct agtggactga agaactggtt tcgtgaagta atgcccggag gctttaccta
     1261 cgatcgtcat attcaatata aaggcgatgg gagtatccat gcaaaacacc aacactttat
     1321 gaaaaatggg acttatcaca acattgtaga atttaccggt caggatttta aagaaaatag
     1381 tccagtctta actggagata tgaatgtctc attaccgaat gaagtcccac aaatacccag
     1441 agatgatgga gtagaatgcc cagtgacctt gctttatcct ttattatcgg ataaatcaaa
     1501 atacgtcgag gctcaccaat atacaatctg caagcctctt cataatcaac cagcacctga
     1561 tgtcccatat cactggattc gtaaacaata cacacaaagc aaagatgatg ccgaggaacg
     1621 cgatcatatc tgtcaatcag agactctcga agcacactta aagggcatgg acgagctgta
     1681 taagcgaccg gttcacgtgg gaagtggggc tacaaacttc tcactgctga aacaggccgg
     1741 ggatgtggag gaaaacccag gaccacacgt gactagtatg gtgagcgagc tgattaagga
     1801 gaacatgccc atgaagctgt acatggaggg caccgtgaac aaccaccact tcaagtgcac
     1861 atccgagggc gaaggcaagc cctacgaggg cacccagacc atgagaatca aggtcgtcga
     1921 gggcggccct ctccccttcg ccttcgacat cctggctacc agcttcatgt acggcagcag
     1981 aaccttcatc aagcaccctc cgggcatccc cgacttcttt aagcagtcct tccctgaggg
     2041 cttcacatgg gagagagtca ccacatacga agacgggggc gtgctgaccg ctacccagga
     2101 caccagcctc caggacggct gcctcatcta caacgtcaag gttagagggg tgaacttccc
     2161 agccaacggc cctgtgatgc agaagaaaac actcggctgg gaggcctcca ccgagacgat
     2221 gtaccctgct gacggaggac tggaaggcgc atgtgacatg gctctgaagc tcgtgggagg
     2281 aggacacctg atctgcaacc ttgagacaac atacagatcc aagaaacccg ctacgaacct
     2341 caagatgccc ggcgtctaca acgtggacca cagactggaa agaatcaagg aggccgacga
     2401 tgagacatac gtcgagcagc acgaggtggc tgtggccaga tactctactg gtggcgctgg
     2461 tgatggaggt aaataagaat tcgatatcaa gcttatcgat aatcaacctc tggattacaa
     2521 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     2581 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     2641 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     2701 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     2761 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     2821 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     2881 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat
     2941 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     3001 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     3061 tcggatctcc ctttgggccg cctccccgca tcgataccgt cgacccgggc ggccgcttcg
     3121 agcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     3181 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     3241 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggagat
     3301 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg ataaggatct
     3361 tcctagagca tggctacgta gataagtagc atggcgggtt aatcattaac tacaaggaac
     3421 ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc
     3481 gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc
     3541 gcagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     3601 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     3661 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     3721 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     3781 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     3841 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     3901 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     3961 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     4021 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     4081 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     4141 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     4201 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     4261 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     4321 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     4381 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     4441 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     4501 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     4561 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     4621 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     4681 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     4741 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     4801 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     4861 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     4921 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     4981 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     5041 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     5101 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     5161 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     5221 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     5281 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     5341 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     5401 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     5461 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     5521 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     5581 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     5641 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     5701 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     5761 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     5821 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     5881 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     5941 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
     6001 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
     6061 cgcgctcgct cgctcactga ggccgcccgg gcaaagcccg ggcgtcgggc gacctttggt
     6121 cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg
     6181 ggttccttgt agttaatgat taacccgcca tgctact
//
