LOCUS       Exported                7281 bp ds-DNA     circular SYN 11-MAY-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7281)
  AUTHORS   Ken Chan
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, May 30, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7281
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    1..130
                     /label=L-ITR
     promoter        144..1243
                     /label=CAG
                     /note="CAG"
     enhancer        201..504
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        506..783
                     /label=chicken beta-actin promoter
     misc_feature    1250..1255
                     /label=Kozak
     CDS             1256..1333
                     /codon_start=1
                     /label=NLS-Element
                     /translation="MSSDDEATADSQHSTPPKKKRKVEDP"
     CDS             1304..1324
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1334..2025
                     /codon_start=1
                     /product="mScarlet"
                     /label=mScarlet
                     /note="red fluorescent protein"
                     /protein_id="APD76535.1"
                     /translation="VSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGGA
                     VTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIKM
                     ALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRHS
                     TGGMDELY"
     misc_feature    2027..2092
                     /label=p2A peptide
                     /note="p2A peptide"
     CDS             2036..2092
                     /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             2093..3741
                     /label=hluc
                     /note="gb_ay535007   derived from firefly; hluc+"
     misc_feature    3748..4339
                     /label=WPRE
     polyA_signal    4346..4578
                     /label=SV40 late polyadenylation signal
     repeat_region   4595..4735
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      4810..5265
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5547..5651
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5652..6461
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in
                     bacteria or G418 (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      6609..7276
                     /direction=RIGHT
                     /label=pUC ori
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac tagttattaa tagtaatcaa ttacggggtc attagttcat
      181 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      241 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      301 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      361 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      421 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      481 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      541 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      601 gcgatggggg cggggggggg gggggggcgc gcgccaggcg gggcggggcg gggcgagggg
      661 cggggcgggg cgaggcggag aggtgcggcg gcagccaatc agagcggcgc gctccgaaag
      721 tttcctttta tggcgaggcg gcggcggcgg cggccctata aaaagcgaag cgcgcggcgg
      781 gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc
      841 ccgccccggc tctgactgac cgcgttactc ccacaggtga gcgggcggga cggcccttct
      901 cctccgggct gtaattagcg cttggtttaa tgacggcttg tttcttttct gtggctgcgt
      961 gaaagccttg aggggctccg ggagcgccgg caggaaggaa atgggcgggg agggccttcg
     1021 tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc cgcgggggga
     1081 cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg tgaccggcgg
     1141 ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc tcctgggcaa
     1201 cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttgacgcgtg ccaccatgtc
     1261 tagtgatgat gaggctactg ctgactctca acattctact cctccaaaaa agaagagaaa
     1321 ggtagaagac cccgtgagca agggcgaggc agtgatcaag gagttcatgc ggttcaaggt
     1381 gcacatggag ggctccatga acggccacga gttcgagatc gagggcgagg gcgagggccg
     1441 cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt
     1501 ctcctgggac atcctgtccc ctcagttcat gtacggctcc agggccttca ccaagcaccc
     1561 cgccgacatc cccgactact ataagcagtc cttccccgag ggcttcaagt gggagcgcgt
     1621 gatgaacttc gaggacggcg gcgccgtgac cgtgacccag gacacctccc tggaggacgg
     1681 caccctgatc tacaaggtga agctccgcgg caccaacttc cctcctgacg gccccgtaat
     1741 gcagaagaag acaatgggct gggaagcgtc caccgagcgg ttgtaccccg aggacggcgt
     1801 gctgaagggc gacattaaga tggccctgcg cctgaaggac ggaggccgct acctggcgga
     1861 cttcaagacc acctacaagg ccaagaagcc cgtgcagatg cccggcgcct acaacgtgga
     1921 ccgcaagttg gacatcacct cccacaacga ggactacacc gtggtggaac agtacgaacg
     1981 ctccgagggc cgccactcca ccggcggcat ggacgagctg tataagggaa gcggagctac
     2041 taacttcagc ctgctgaagc aggctggaga cgtggaggag aaccctgggc ccgaggatgc
     2101 caagaatatt aagaaaggcc ctgccccatt ctaccctctg gaagatggca ctgctggtga
     2161 gcaactgcac aaggccatga agaggtatgc cctggtccct ggcaccattg ccttcactga
     2221 tgctcacatt gaggtggaca tcacctatgc tgaatacttt gagatgtctg tgaggctggc
     2281 agaagccatg aaaagatatg gactgaacac caaccacagg attgtggtgt gctctgagaa
     2341 ctctctccag ttcttcatgc ctgtgttagg agccctgttc attggagtgg ctgtggcccc
     2401 tgccaatgac atctacaatg agagagagct cctgaacagc atgggcatca gccagccaac
     2461 tgtggtcttt gtgagcaaga agggcctgca aaagatcctg aatgtgcaga agaagctgcc
     2521 catcatccag aagatcatca tcatggacag caagactgac taccagggct tccagagcat
     2581 gtataccttt gtgaccagcc acttaccccc tggcttcaat gagtatgact ttgtgcctga
     2641 gagctttgac agggacaaga ccattgctct gattatgaac agctctggct ccactggact
     2701 gcccaaaggt gtggctctgc cccacagaac tgcttgtgtg agattcagcc atgccagaga
     2761 ccccatcttt ggcaaccaga tcatccctga cactgccatc ctgtctgtgg ttccattcca
     2821 tcatggcttt ggcatgttca caacactggg gtacctgatc tgtggcttca gagtggtgct
     2881 gatgtatagg tttgaggagg agctgtttct gaggagccta caagactaca agatccagtc
     2941 tgccctgctg gtgcccactc tgttcagctt ctttgccaag agcaccctca ttgacaagta
     3001 tgacctgagc aacctgcatg agattgcctc tggaggagca cccctgagca aggaggtggg
     3061 tgaggctgtg gcaaagaggt tccatctccc aggaatcaga cagggctatg gcctgactga
     3121 gaccacctct gccatcctca tcacccctga aggagatgac aagcctggtg ctgtgggcaa
     3181 ggtggttccc ttttttgagg ccaaggtggt ggacctggac actggcaaga ccctgggagt
     3241 gaaccagagg ggtgagctgt gtgtgagggg tcccatgatc atgtctggct atgtgaacaa
     3301 ccctgaggcc accaatgccc tgattgacaa ggatggctgg ctgcactctg gtgacattgc
     3361 ctactgggat gaggatgagc actttttcat tgtggacagg ctgaagagcc tcatcaagta
     3421 caaaggctac caagtggcac ctgctgagct agagagcatc ctgctccagc accccaacat
     3481 ctttgatgct ggtgtggctg gcctgcctga tgatgatgct ggagagctgc ctgctgctgt
     3541 tgtggttctg gagcatggaa agaccatgac tgagaaggag attgtggact atgtggccag
     3601 tcaggtgacc actgccaaga agctgagggg aggtgtggtg tttgtggatg aggtgccaaa
     3661 gggtctgact ggcaagctgg atgccagaaa gatcagagag atcctgatca aggccaagaa
     3721 gggtggcaaa atcgccgtct agaattctaa tcaacctctg gattacaaaa tttgtgaaag
     3781 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     3841 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     3901 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     3961 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     4021 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     4081 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     4141 gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     4201 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     4261 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     4321 ttgggccgcc tccccgcctc tcgagttcga gcagacatga taagatacat tgatgagttt
     4381 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     4441 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     4501 cattttatgt ttcaggttca gggggagatg tgggaggttt tttaaagcaa gtaaaacctc
     4561 tacaaatgtg gtaaaatccg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc
     4621 actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc
     4681 ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc
     4741 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc
     4801 aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     4861 gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct
     4921 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     4981 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac
     5041 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     5101 ttaatagtgg actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt
     5161 ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac
     5221 aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca
     5281 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     5341 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     5401 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     5461 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     5521 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     5581 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     5641 aaaggaagag tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca
     5701 tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga
     5761 caatctatcg cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag
     5821 gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta
     5881 tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca
     5941 ctgcgatccc cggaaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa
     6001 atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt
     6061 gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg
     6121 gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct
     6181 ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt
     6241 tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac
     6301 gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt
     6361 tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga
     6421 ataaattgca gtttcatttg atgctcgatg agtttttcta actgtcagac caagtttact
     6481 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     6541 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     6601 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     6661 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     6721 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     6781 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     6841 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     6901 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     6961 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     7021 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     7081 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     7141 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     7201 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     7261 gctggccttt tgctcacatg t
//
