LOCUS       Exported                8771 bp ds-DNA     circular SYN 03-DEC-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8771)
  AUTHORS   Triple Threat
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Oct 9, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8771
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      30..618
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        686..926
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     enhancer        1128..1413
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        1415..1692
                     /label=chicken beta-actin promoter
     intron          1693..1921
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     misc_feature    1911..1921
                     /label=Hyrid Intron
     CDS             1939..1995
                     /codon_start=1
                     /label=BP-NLS
                     /translation="MKRTADGSEFESPKKKRKV"
     CDS             4789..4809
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T 
                     antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    4843..5050
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        5119..5422
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5423..5626
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     misc_feature    5655..5663
                     /label=Kozak
     CDS             5661..6371
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     DDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     rep_origin      6928..7383
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        7665..7769
                     /gene="bla"
                     /label=AmpR promoter
     CDS             7770..8630
                     /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"
ORIGIN
        1 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
       61 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
      121 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
      181 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
      241 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
      301 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
      361 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
      421 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
      481 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
      541 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
      601 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
      661 tggccttttg ctcacatgct ctagagaggg cctatttccc atgattcctt catatttgca
      721 tatacgatac aaggctgtta gagagataat tggaattaat ttgactgtaa acacaaagat
      781 attagtacaa aatacgtgac gtagaaagta ataatttctt gggtagtttg cagttttaaa
      841 attatgtttt aaaatggact atcatatgct taccgtaact tgaaagtatt tcgatttctt
      901 ggctttatat atcttgtgga aaggacgaaa caccggggtc ttcgagaaga cctggctcgt
      961 ccaactgcgg ttgaacgagc acaggctgag acattcgtac gcaccctggg atttccccag
     1021 tcccaggaac tgcatagcgg atgccagttg atggagcaat ctatcagata agccaggggg
     1081 aacaatcacc tctctgtatc agagagagtt ttactttttt tggtacccgt tacataactt
     1141 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaatagta
     1201 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
     1261 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
     1321 aaatggcccg cctggcattg tgcccagtac atgaccttat gggactttcc tacttggcag
     1381 tacatctacg tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc
     1441 actctcccca tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta
     1501 ttttgtgcag cgatgggggc gggggggggg ggggggcgcg cgccaggcgg ggcggggcgg
     1561 ggcgaggggc ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg
     1621 ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc
     1681 gcgcggcggg cgggagtcgc tgcgacgctg ccttcgcccc gtgccccgct ccgccgccgc
     1741 ctcgcgccgc ccgccccggc tctgactgac cgcgttactc ccacaggtga gcgggcggga
     1801 cggcccttct cctccgggct gtaattagct gagcaagagg taagggttta agggatggtt
     1861 ggttggtggg gtattaatgt ttaattacct ggagcacctg cctgaaatca ctttttttca
     1921 ggttggaccg gtgccaccat gaaacggaca gccgacggaa gcgagttcga gtcaccaaag
     1981 aagaagcgga aagtcatgga ggcatctcca gcaagcggac caaggcacct gatggacccc
     2041 cacatcttca cctctaactt taacaatggc atcggcaggc acaagacata cctgtgctat
     2101 gaggtggagc gcctggacaa tggcaccagc gtgaagatgg atcagcacag aggcttcctg
     2161 cacaaccagg ccaagaatct gctgtgcggc ttctacggcc ggcacgcaga gctgagattt
     2221 ctggacctgg tgcctagcct gcagctggat ccagcccaga tctatagggt gacctggttc
     2281 atcagctact ccccatgctt ttccgcggga tgtgcaggag aggtgcgcgc cttcctgcag
     2341 gagaacacac acgtgcggct gagaatcttt gccgcccgga tcttcgacta tgatcctctg
     2401 tacaaggagg ccctgcagat gctgagagac gcaggagccc aggtgtccat catgacctat
     2461 gatgagttca agcactgctg ggacacattt gtggatcacc agggctgtcc ctttcagcct
     2521 tgggacggac tggatgagca ctcccaggcc ctgtctggca ggctgagggc catcctgcag
     2581 aaccagggca atagcggcag cgagactccc gggacctcag agtccgccac acccgaaagt
     2641 atgatggccg tggtgtacgt gatcagcaag tctggcaaac ccctgatgcc caccacccgg
     2701 tgcggccacg tgagaatcct gctgaaggag ggcaaggcca gagtggtgga gcggaaaccg
     2761 ttcaccatcc agctgaccta cgagagcgcg gaggaaacac agcctctggt cctgggcatt
     2821 gctcctggac gcaccaatat cggcatgagc gtggtgacag aatccggaga gagcgtgttc
     2881 aacgcccaga tcagaaccag aaacaaggac gtgcctaagc tgatgaaaga cagaaagcag
     2941 taccggatgg ctcacagacg gctgaagcgg cgctgcaagc gaagaagaag agccaaggcc
     3001 gccggcaccg cattcgagga aggagaaaag cagagactgc tgcctggctg tttcaagcct
     3061 atcacatgta agagcatcag aaataaggaa gctagattca ataacaggaa gcggcctgtg
     3121 ggatggctga ccccaaccgc caaccacctg ctggtgaccc acctgaacgt ggtcaagaag
     3181 gtgcaaaaga tcctgcctgt ggccaaggtg gtgctggaac tgaatcggtt cagcttcatg
     3241 gccatgaaca accccaaggt gcagcggtgg cagtaccaga gaggccctct gtacggtaaa
     3301 ggcagcgtgg aagaggccgt gagcatgcag caggacggcc actgcctgtt ttgcaagcac
     3361 ggcattgacc actatcacca tgtggttcct aggagaaaga acggctctga aaccctggag
     3421 aaccgggtgg gcctgtgcga ggaacaccac cggctcgtcc acacagataa ggagtgggag
     3481 gccaatctgg cttctaagaa gagcggaatg aacaagaaat accacgccct gagcgtgctg
     3541 aaccagatca tcccatacct ggccgaccag cttgccgata tgttccccgg caacttctgt
     3601 gtgaccagcg gacaggatac atacctgttt cgggaagaac atggcatccc caaggaccac
     3661 tacctggatg cctactgcat cgcctgcagc gccctgaccg acgccaagaa ggtgagcagc
     3721 cctaaaggca gaccttacat ggtgagacag ttcagacgcc acgatagaca ggcctgtcac
     3781 aaggccaacc tgaacagaag atactacatg ggcggcaaac tggttgctac aaacagacat
     3841 aaggccatgg accagaaaac cgacagcctg gaggagtaca gagccgccca cagcgccgct
     3901 gacgtatcca agctgacagt gaagcaccct tctgctcaat ataaggacat gtcccggatc
     3961 atgcctggca gcatcctcgt gtccggcgag ggcaagctct ttacactgag aagatccgag
     4021 ggcagaaaca agggccaagt gaactacttc gtgagcaccg agggcatcaa gtactgggcc
     4081 agaaagtgcc agtatctgag aaacaatggc ggcctgcaga tctacgtgaa aaggccggcg
     4141 gccacgaaga aggccggcca ggcaaagaag aaaaagagcg gcgggagcgg cgggagcggg
     4201 gggagcacta atctgtcaga catcatcgag aaggagacag gcaagcagct ggtcatccag
     4261 gagagcatcc tgatgctgcc agaggaggtg gaggaagtga tcggcaacaa gcccgagtcc
     4321 gatatcctgg tgcacaccgc ctacgacgag agcacagatg agaatgtgat gctgctgacc
     4381 tccgacgccc ccgagtataa gccttgggcc ctggtcatcc aggattctaa cggcgagaat
     4441 aagatcaaga tgctgagcgg aggatccgga ggatctggag gcagcaccaa cctgtctgac
     4501 atcatcgaga aggagacagg caagcagctg gtcatccagg agagcatcct gatgctgcca
     4561 gaggaggtgg aggaagtgat cggcaacaag cccgagtccg atatcctggt gcacaccgcc
     4621 tacgacgaga gcacagatga gaatgtgatg ctgctgacct ccgacgcccc cgagtataag
     4681 ccttgggccc tggtcatcca ggattctaac ggcgagaata agatcaagat gctgtctggt
     4741 ggttcttctg gcggctcaaa gagaaccgcc gacggcagcg aatccgagcc caagaagaag
     4801 aggaaagtct aagaattcct agagctcgct gatcagcctc gactgtgcct tctagttgcc
     4861 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     4921 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     4981 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagag aatagcaggc
     5041 atgctgggga gcggccgcta gttattaata gtaatcaatt acggggtcat tagttcatag
     5101 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     5161 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     5221 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     5281 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
     5341 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
     5401 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
     5461 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
     5521 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
     5581 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg
     5641 tcagatccgc tagcgccacc atggtgagca agggcgagga ggataacatg gccatcatca
     5701 aggagttcat gcgcttcaag gtgcacatgg agggctccgt gaacggccac gagttcgaga
     5761 tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga
     5821 ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc atgtacggct
     5881 ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttgaagctg tccttccccg
     5941 agggcttcaa gtgggagcgc gtgatgaact tcgatgacgg cggcgtggtg accgtgaccc
     6001 aggactcctc cctgcaggac ggcgagttca tctacaaggt gaagctgcgc ggcaccaact
     6061 tcccctccga cggccccgta atgcagaaga aaaccatggg ctgggaggcc tcctccgagc
     6121 ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa gcagaggctg aagctgaagg
     6181 acggcggcca ctacgacgct gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc
     6241 tgcccggcgc ctacaacgtc aacatcaagt tggacatcac ctcccacaac gaggactaca
     6301 ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc atggacgagc
     6361 tgtataagtg atgtacaccg ggatgcagaa attgatgatc tattaaacaa taaagatgtc
     6421 cactaaaatg gaagtttttc ctgtcatact ttgttaagaa gggtgagaac agagtaccta
     6481 cattttgaat ggaaggattg gagctacggg ggtgggggtg gggtgggatt agataaatgc
     6541 ctgctcttta ctgaaggctc tttactattg ctttatgata atgtttcata gttggatatc
     6601 ataatttaaa caagcaaaac caaattaagg gccagctcat tcctcccact catgatctat
     6661 agatctatag atctctcgtg ggatcattgt ttttctcttg attcccactt tgtggttcta
     6721 agtactgtgg tttccaaatg tgtcagtttc atagcctgaa gaacgagatc agcagcctct
     6781 gttccacata cacttcattc tcagtattgt tttgccaagt tctaattcca tcagaagctg
     6841 gtcgacctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     6901 caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg
     6961 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     7021 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     7081 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     7141 atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     7201 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     7261 ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     7321 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa
     7381 ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
     7441 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     7501 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     7561 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     7621 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     7681 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     7741 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     7801 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     7861 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     7921 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     7981 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     8041 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     8101 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     8161 ctgcggccaa cttacttctg acaacgattg gaggaccgaa ggagctaacc gcttttttgc
     8221 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     8281 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     8341 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     8401 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     8461 ataaatctgg agccggtgag cgtggaagcc gcggtatcat tgcagcactg gggccagatg
     8521 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     8581 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     8641 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     8701 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     8761 actgagcgtc a
//
