LOCUS       Exported                8639 bp ds-DNA    circular SYN 20-9ÔÂ-2025
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    ICBE
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8639)
  AUTHORS   Triple Threat
  TITLE     Direct Submission
  JOURNAL   Exported 2025Äê9ÔÂ20ÈÕ from SnapGene 2.3.2
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8639
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      30..618
                     /direction=RIGHT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    638..923
                     /note="CMV enhancer"
     misc_feature    717
                     /note="A-to-G R2M4"
     promoter        925..1202
                     /note="chicken beta-actin promoter"
     intron          1203..1431
                     /note="hybrid intron"
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     misc_feature    1421..1431
                     /note="Hyrid Intron"
     CDS             1452..2819
                     /codon_start=1
                     /note="enDelIscB(D60A)"
                     /translation="SRVLVVDADRRPLAPCTPRRARLLLNSGKAAVLRRYPFTIILKQS
                     YPTASPRPVRLKLAPGSKTTGIAVVTEATGEVVWAAELQHRGQLIKNALEKRRALRRSR
                     RNRKTRYRPARFLNRKRTGPPLLSSADTVSTLGKWLPPSLQHRIEVIMTWVHRLRRYLP
                     ITAISQEIVRFDMQKMQNPEISGVEYQQGTLFGYEVREYLLDKWRRQCGYCGAKDKRLE
                     VDHIVPRSHGGSDRVSNLTLSCEPCNKRKNQRPAAVFLAKKPEVLQKLQRQAKAPLKDA
                     AAVNSTRYALLERLKATGLPVEVASGGRTKFNRSRRQIPKTHWLDAACVGASTPEVLKW
                     EAVRPLAIKAMGHGKRQVVNVDAYGFPRGKAKGIPVHPFRTGDIVRAEIPKGKYKGTYV
                     SRIAETTTSKPLAGFKSKTGKRIQCHTKHMTKLFNRDGYGYGFLKAPEPRQTVISES"
     variation       1482..1484
                     /note="C12R"
     variation       1626..1628
                     /note="D60A"
     variation       1737..1739
                     /note="S97K"
     variation       1746..1748
                     /note="S100A"
     variation       1758..1760
                     /note="G104S"
     variation       1797..1799
                     /note="W117F"
     misc_feature    1872..1874
                     /note="A142P"
     misc_feature    2397..2399
                     /note="E317R"
     variation       2466..2468
                     /note="Q340K"
     variation       2634..2636
                     /note="V396K"
     variation       2754..2756
                     /note="S436R"
     CDS             2820..2867
                     /codon_start=1
                     /note="xten"
                     /translation="SGSETPGTSESATPES"
     CDS             2868..3464
                     /codon_start=1
                     /product="APOBEC3A (A3A) "
                     /note="A3A(BEACON2)"
                     /translation="MEASPASGPRHLMDPHIFTSNFNNGIGRHKTYLCYEVERLDNGTS
                     VKMDQHRGFLHNQAKNLLCGFYGRHAELRFLDLVPSLQLDPAQIYRVTWFISYSPCFSA
                     GCAGEVRAFLQENTHVRLRIFAARIFDYDPLYKEALQMLRDAGAQVSIMTYDEFKHCWD
                     TFVDHQGCPFQPWDGLDEHSQALSGRLRAILQNQGN"
     misc_feature    3177..3179
                     /note="W104A"
     CDS             3465..3512
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /note="nucleoplasmin NLS"
                     /translation="KRPAATKKAGQAKKKK"
     misc_feature    3513..3542
                     /note="10AA linker"
     CDS             3543..3791
                     /codon_start=1
                     /note="UGI"
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             3822..4070
                     /codon_start=1
                     /locus_tag="
                     "
                     /old_locus_tag="
                     "
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /note="UGI"
                     /note="
                     "
                     /protein_id="
                     "
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             4125..4145
                     /codon_start=1
                     /locus_tag="
                     "
                     /product="nuclear localization signal of SV40 large T 
                     antigen"
                     /note="SV40 NLS"
                     /note="
                     "
                     /protein_id="
                     "
                     /translation="PKKKRKV"
     misc_feature    4173..4177
                     /note="i16-2"
     polyA_signal    4179..4354
                     /note="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    4355..4637
                     /note="F3"
     promoter        4387..4636
                     /label="U6 Promoter"
                     /note="U6 promoter"
                     /note="RNA polymerase III promoter for human U6 snRNA"
     misc_feature    4637
                     /note="Bsa1"
     misc_feature    4637
                     /note="replace"
     misc_feature    4638..4920
                     /note="F4"
     misc_feature    4638..4658
                     /note="Bsa1"
     misc_feature    4638..4656
                     /note="replace"
     misc_feature    4659..4906
                     /note="Delta-35-41+GAAA+T205A+Delta-221-240+GAAA"
     misc_feature    4659..4677
                     /note="DR"
     misc_feature    4678..4681
                     /note="LOOP"
     misc_feature    4682..4712
                     /note="anti-repeat"
     misc_feature    4694..4699
                     /note="Á¬Ðø6T"
     misc_feature    4860
                     /note="T205A"
     enhancer        4987..5290
                     /note="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5291..5494
                     /note="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     misc_feature    5523..5531
                     /note="Kozak"
     CDS             5529..6239
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /note="mCherry"
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     DDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     rep_origin      6796..7251
                     /direction=RIGHT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        7533..7637
                     /gene="bla"
                     /note="AmpR promoter"
     CDS             7638..8498
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="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 aggtacccgt tacataactt acggtaaatg
      661 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaatagta acgccaatag
      721 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      781 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      841 cctggcattg tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      901 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
      961 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
     1021 cgatgggggc gggggggggg ggggggcgcg cgccaggcgg ggcggggcgg ggcgaggggc
     1081 ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
     1141 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
     1201 cgggagtcgc tgcgacgctg ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc
     1261 ccgccccggc tctgactgac cgcgttactc ccacaggtga gcgggcggga cggcccttct
     1321 cctccgggct gtaattagct gagcaagagg taagggttta agggatggtt ggttggtggg
     1381 gtattaatgt ttaattacct ggagcacctg cctgaaatca ctttttttca ggttggaccg
     1441 gtgccaccat gagcagagtg ctggtggtgg atgctgatag gcgccctctg gccccttgca
     1501 cccctcggag agccagactg ctgctgaatt ctggcaaagc tgccgtgctg agacggtacc
     1561 ctttcaccat catcctgaag cagagctacc ccaccgccag ccctagaccc gtgcggctga
     1621 agctggcccc cggaagcaag accaccggca tcgccgtggt gacagaggcc acaggagaag
     1681 tggtgtgggc cgctgagctg cagcacagag gccagctgat caagaacgcc ctggaaaaga
     1741 gaagagccct gcggcgcagc cggcgaaacc ggaagacacg gtatagacct gccagattcc
     1801 tgaatagaaa gagaacagga cctccactgc tgagcagcgc cgacactgtg tctaccctgg
     1861 gcaagtggtt gcccccgagc ctgcagcata ggatcgaggt gatcatgacc tgggtccaca
     1921 gactgagaag atacctgccc atcaccgcca tttctcagga gatcgtgaga ttcgacatgc
     1981 aaaagatgca gaaccccgaa atcagcggcg tggaatacca gcagggcacc ctcttcggct
     2041 acgaggtgcg ggaatacctc ctggacaagt ggcggagaca atgtggctac tgcggcgcta
     2101 aggacaagag acttgaggtc gaccacatcg ttcctcggag ccacggcggc agcgacagag
     2161 tgtccaacct gacactgtcc tgcgagcctt gcaacaaaag aaagaaccag aggccagctg
     2221 ctgtgttcct ggctaaaaag cccgaggtcc tgcagaagct gcagcggcag gccaaggccc
     2281 ctctgaagga cgccgccgcc gtgaacagca cccgttacgc cctgctggaa agactgaagg
     2341 ccaccggcct gcctgtggaa gtggccagcg gcggaagaac caagttcaat agaagcagga
     2401 gacagatccc caagacccac tggctggatg ccgcctgcgt gggcgcttct acacctgagg
     2461 tgctgaagtg ggaggccgtt agaccactgg ccatcaaggc catgggccac ggcaaacggc
     2521 aggtggtcaa cgtggacgcc tacggatttc ctcggggcaa ggccaaaggc atccctgtgc
     2581 accccttcag aaccggagat atcgtgcggg ccgagattcc aaagggcaaa tacaagggaa
     2641 catacgtgtc cagaatcgcc gagacaacca catctaagcc tctagcaggc tttaagagca
     2701 agaccggcaa gcggatccag tgtcacacca agcacatgac aaagctgttc aaccgcgacg
     2761 gctatggcta cggcttcctg aaagcccctg aacctagaca gaccgtgatc agcgagtcta
     2821 gcggcagcga gactcccggg acctcagagt ccgccacacc cgaaagtatg gaggcatctc
     2881 cagcaagcgg accaaggcac ctgatggacc cccacatctt cacctctaac tttaacaatg
     2941 gcatcggcag gcacaagaca tacctgtgct atgaggtgga gcgcctggac aatggcacca
     3001 gcgtgaagat ggatcagcac agaggcttcc tgcacaacca ggccaagaat ctgctgtgcg
     3061 gcttctacgg ccggcacgca gagctgagat ttctggacct ggtgcctagc ctgcagctgg
     3121 atccagccca gatctatagg gtgacctggt tcatcagcta ctccccatgc ttttccgcgg
     3181 gatgtgcagg agaggtgcgc gccttcctgc aggagaacac acacgtgcgg ctgagaatct
     3241 ttgccgcccg gatcttcgac tatgatcctc tgtacaagga ggccctgcag atgctgagag
     3301 acgcaggagc ccaggtgtcc atcatgacct atgatgagtt caagcactgc tgggacacat
     3361 ttgtggatca ccagggctgt ccctttcagc cttgggacgg actggatgag cactcccagg
     3421 ccctgtctgg caggctgagg gccatcctgc agaaccaggg caataaaagg ccggcggcca
     3481 cgaagaaggc cggccaggca aagaagaaaa agagcggcgg gagcggcggg agcgggggga
     3541 gcactaatct gtcagatatt attgaaaagg aaaccggtaa gcaactggtt atccaggaat
     3601 ccatcctcat gctcccagag gaggtggaag aagtcattgg gaacaagccg gaaagcgata
     3661 tactcgtgca caccgcctac gacgagagca ccgacgagaa tgtcatgctt ctgactagcg
     3721 acgcccctga atacaagcct tgggctctgg tcatacagga tagcaacggt gagaacaaga
     3781 ttaagatgct cagcggagga tccggaggat ctggaggcag caccaacctg tctgacatca
     3841 tcgagaagga gacaggcaag cagctggtca tccaggagag catcctgatg ctgccagagg
     3901 aggtggagga agtgatcggc aacaagcccg agtccgatat cctggtgcac accgcctacg
     3961 acgagagcac agatgagaat gtgatgctgc tgacctccga cgcccccgag tataagcctt
     4021 gggccctggt catccaggat tctaacggcg agaataagat caagatgctg tctggtggtt
     4081 cttctggcgg ctcaaagaga accgccgacg gcagcgaatc cgagcccaag aagaagagga
     4141 aagtctaaga attcctagag ctcgctgatc agcctcgact gtgccttcta gttgccagcc
     4201 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     4261 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     4321 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagagaata gcaggcatgc
     4381 tggggagagg gcctatttcc catgattcct tcatatttgc atatacgata caaggctgtt
     4441 agagagataa ttggaattaa tttgactgta aacacaaaga tattagtaca aaatacgtga
     4501 cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt taaaatggac
     4561 tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata tatcttgtgg
     4621 aaaggacgaa acaccgggga gaccgagaga gggtctcagt tgtagcctcc actcattgaa
     4681 aaatgagtgc gggaaaccgc agctacaacc agactaagtt ctttgggaac tacgttattg
     4741 tcgtcatcat acccacgggt gcgtgccagc ctgtggctct atggtcgtgc attaaacagg
     4801 gactttggag ggtgcccagt gtgtacggcg tcaaaagcgg cgataacatt gtcgaggcaa
     4861 actttacgtg acggcgaaag cccgatcgct agctttaagg ctaatttttt atttttttgc
     4921 ggccgctagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
     4981 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     5041 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     5101 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     5161 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     5221 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     5281 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
     5341 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
     5401 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
     5461 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta
     5521 gcgccaccat ggtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc
     5581 gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg
     5641 gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc
     5701 ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg
     5761 tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt
     5821 gggagcgcgt gatgaacttc gatgacggcg gcgtggtgac cgtgacccag gactcctccc
     5881 tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg
     5941 gccccgtaat gcagaagaaa accatgggct gggaggcctc ctccgagcgg atgtaccccg
     6001 aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact
     6061 acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct
     6121 acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac
     6181 agtacgaacg cgccgagggc cgccactcca ccggcggcat ggacgagctg tataagtgat
     6241 gtacaccggg atgcagaaat tgatgatcta ttaaacaata aagatgtcca ctaaaatgga
     6301 agtttttcct gtcatacttt gttaagaagg gtgagaacag agtacctaca ttttgaatgg
     6361 aaggattgga gctacggggg tgggggtggg gtgggattag ataaatgcct gctctttact
     6421 gaaggctctt tactattgct ttatgataat gtttcatagt tggatatcat aatttaaaca
     6481 agcaaaacca aattaagggc cagctcattc ctcccactca tgatctatag atctatagat
     6541 ctctcgtggg atcattgttt ttctcttgat tcccactttg tggttctaag tactgtggtt
     6601 tccaaatgtg tcagtttcat agcctgaaga acgagatcag cagcctctgt tccacataca
     6661 cttcattctc agtattgttt tgccaagttc taattccatc agaagctggt cgacctgcag
     6721 gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt
     6781 caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     6841 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     6901 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     6961 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg
     7021 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     7081 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcgggct
     7141 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     7201 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca
     7261 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
     7321 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     7381 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
     7441 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     7501 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     7561 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     7621 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     7681 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     7741 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     7801 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     7861 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     7921 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     7981 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     8041 tacttctgac aacgattgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     8101 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     8161 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     8221 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     8281 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     8341 ccggtgagcg tggaagccgc ggtatcattg cagcactggg gccagatggt aagccctccc
     8401 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     8461 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     8521 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     8581 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtca
//
