LOCUS       Exported                6223 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 6223)
  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..6223
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        279..556
                     /label=chicken beta-actin promoter
     misc_feature    977..982
                     /label=Kozak sequence
     CDS             983..1690
                     /codon_start=1
                     /label=(n1)mStayGold(E138D)(c4）
                     /translation="MASTGEELFTGVVPFKFQLKGTINGKSFTVEGEGEGNSHEGSHKG
                     KYVCTSGKLPMSWAALGTSFGYGMKYYTKYPSGLKNWFHEVMPEGFTYDRHIQYKGDGS
                     IHAKHQHFMKNGTYHNIVEFTGQDFKENSPVLTGDMNVSLPNDVQHIPRDDGVECPVTL
                     LYPLLSDKSKCVEAHQNTICKPLHNQPAPDVPYHWIRKQYTQSKDDTEERDHICQSETL
                     EAHLPWHEPSASAV"
     misc_feature    995..1021
                     /label=n1
     primer_bind     1402..1457
                     /label=0707-2-E138D-F
     misc_feature    1421..1423
                     /label=E138D
     misc_feature    1661..1690
                     /label=c4
     primer_bind     complement(1671..1713)
                     /label=0718-QC2-6-R
     CDS             1715..1771
                     /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             1784..2479
                     /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(2459..2503)
                     /label=0714-FusionRed-R
     misc_feature    2507..3095
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(2978..2989)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3138..3259
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   3421..3550
                     /label=AAV2 ITR
     rep_origin      complement(3787..4375)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4546..5406)
                     /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(5407..5511)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5537..5992)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     repeat_region   6064..6193
                     /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 tacaggggag gagctgttca ccggggtggt
     1021 gccatttaaa tttcaactta aaggaaccat caatggcaaa tcgtttaccg ttgaaggcga
     1081 aggtgaagga aattcacatg aaggttctca taaaggaaaa tatgtttgta caagtggaaa
     1141 actaccgatg tcatgggcag cacttggaac atcctttggt tatggaatga aatattatac
     1201 caaatatcct agtggactga agaactggtt tcatgaagta atgcctgaag gctttaccta
     1261 cgatcgtcat attcaatata aaggcgatgg gagtatccat gcaaaacacc aacactttat
     1321 gaaaaatggg acttatcaca acattgttga atttactggt caggatttta aagaaaatag
     1381 tccagtctta actggagata tgaatgtctc attaccgaat gacgttcaac atatacccag
     1441 agatgatgga gtagaatgcc cagtgacctt gctttatcct ttattatcgg ataaatcaaa
     1501 atgcgttgag gctcaccaaa atacaatctg caagcctctt cataatcaac cagcacctga
     1561 tgtcccatat cactggattc gtaaacaata cacacaaagc aaagatgata ccgaggaacg
     1621 tgatcatatt tgtcaatcag agactctcga agcacactta ccatggcacg agccttctgc
     1681 ttctgccgtg cgaccggttc acgtgggaag tggggctaca aacttctcac tgctgaaaca
     1741 ggccggggat gtggaggaaa acccaggacc acacgtgact agtatggtga gcgagctgat
     1801 taaggagaac atgcccatga agctgtacat ggagggcacc gtgaacaacc accacttcaa
     1861 gtgcacatcc gagggcgaag gcaagcccta cgagggcacc cagaccatga gaatcaaggt
     1921 cgtcgagggc ggccctctcc ccttcgcctt cgacatcctg gctaccagct tcatgtacgg
     1981 cagcagaacc ttcatcaagc accctccggg catccccgac ttctttaagc agtccttccc
     2041 tgagggcttc acatgggaga gagtcaccac atacgaagac gggggcgtgc tgaccgctac
     2101 ccaggacacc agcctccagg acggctgcct catctacaac gtcaaggtta gaggggtgaa
     2161 cttcccagcc aacggccctg tgatgcagaa gaaaacactc ggctgggagg cctccaccga
     2221 gacgatgtac cctgctgacg gaggactgga aggcgcatgt gacatggctc tgaagctcgt
     2281 gggaggagga cacctgatct gcaaccttga gacaacatac agatccaaga aacccgctac
     2341 gaacctcaag atgcccggcg tctacaacgt ggaccacaga ctggaaagaa tcaaggaggc
     2401 cgacgatgag acatacgtcg agcagcacga ggtggctgtg gccagatact ctactggtgg
     2461 cgctggtgat ggaggtaaat aagaattcga tatcaagctt atcgataatc aacctctgga
     2521 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     2581 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     2641 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     2701 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     2761 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     2821 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     2881 ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac
     2941 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     3001 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     3061 gacgagtcgg atctcccttt gggccgcctc cccgcatcga taccgtcgac ccgggcggcc
     3121 gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     3181 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     3241 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     3301 ggagatgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta aaatcgataa
     3361 ggatcttcct agagcatggc tacgtagata agtagcatgg cgggttaatc attaactaca
     3421 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
     3481 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc
     3541 gagcgcgcag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     3601 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     3661 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     3721 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     3781 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     3841 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     3901 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     3961 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4021 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4081 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4141 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4201 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     4261 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     4321 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     4381 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     4441 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     4501 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     4561 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     4621 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     4681 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     4741 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     4801 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     4861 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     4921 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     4981 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     5041 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     5101 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     5161 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     5221 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     5281 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     5341 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     5401 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     5461 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     5521 ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     5581 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     5641 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     5701 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     5761 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     5821 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     5881 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     5941 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg
     6001 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     6061 cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc
     6121 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc
     6181 actaggggtt ccttgtagtt aatgattaac ccgccatgct act
//
