LOCUS       Exported                7009 bp ds-DNA     circular SYN 04-DEC-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7009)
  AUTHORS   SnapGene License
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Nov 11, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7009
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        complement(24..41)
                     /label=SP6 promoter (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=94.7"
                     /note="/other=promoter"
     misc_feature    53..638
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             639..698
                     /codon_start=1
                     /label=GAP43 N-term
                     /translation="MLCCMRRTKQVEKNDEDQKI"
     CDS             699..1418
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    1569..1624
                     /label=beta-globin poly(A) signal
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=polyA_signal"
     primer_bind     complement(1985..2001)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2009..2025)
                     /label=lac operator
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=protein_bind"
     promoter        complement(2033..2063)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2078..2099
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     protein_bind    complement(2078..2099)
                     /label=CAP binding site
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=protein_bind"
     CDS             complement(2112..2151)
                     /label=lacI (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=3.7"
                     /note="/other=CDS"
     promoter        2158..2353
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2204..2339
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2359..2493
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(2495..2549)
                     /label=lacI (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=5.1"
                     /note="/other=CDS"
     rep_origin      complement(2732..3320)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3491..4351)
                     /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(4352..4456)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        4487..4866
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4868..5146
                     /label=chicken beta-actin promoter
     intron          5147..6163
                     /label=chimeric intron
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=99.9"
                     /note="/match_length=100.0"
                     /note="/other=intron"
     intron          5240..6163
                     /label=chicken b-actin intron + rabbit b-globin intron
     misc_feature    6087..6109
                     /label=pCAGGS FWDSeq
     intron          6164..6216
                     /label=rabbit B-globin intron
     protein_bind    complement(6229..6253)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    6239..6255
                     /label=attL1(1)
                     /note="ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 
                     0} width 5 offset 0
                     ApEinfo_revcolor=#ff66ff
                     ApEinfo_label=attL1
                     ApEinfo_fwdcolor=#ff66ff"
     transit_peptide 6261..6287
                     /label=NLS
                     /note="ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 
                     0} width 5 offset 0
                     ApEinfo_revcolor=cyan
                     ApEinfo_fwdcolor=cyan"
     CDS             6288..6999
                     /codon_start=1
                     /label=mCherry CBC
                     /note="ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 
                     0} width 5 offset 0
                     ApEinfo_revcolor=#ff0080
                     ApEinfo_fwdcolor=#ff0080"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGREFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     misc_feature    7000..7009
                     /label=AttL2
                     /note="ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 
                     0} width 5 offset 0
                     ApEinfo_revcolor=#ff0080
                     ApEinfo_fwdcolor=#ff0080"
ORIGIN
        1 cttgtacaaa gtggttcgat atcctatagt gtcacctaaa taattccgcc cccccctctc
       61 cctccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt
      121 ctatatgtta ttttccacca tattgccgtc ttttggcaat gtgagggccc ggaaacctgg
      181 ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag gaatgcaagg
      241 tctgttgaat gtcgtgaagg aagcagttcc tctggaagct tcttgaagac aaacaacgtc
      301 tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc tctgcggcca
      361 aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag
      421 ttggatagtt gtggaaagag tcaaatggct ctcctcaagc gtattcaaca aggggctgaa
      481 ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt gcacatgctt
      541 tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt
      601 tttcctttga aaaacacgat gataatatgg ccacaaccat gctgtgctgt atgagaagaa
      661 ccaaacaggt tgaaaagaat gatgaggacc aaaagatcat ggtgagcaag ggcgaggagc
      721 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
      781 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
      841 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
      901 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
      961 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
     1021 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     1081 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     1141 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggcg aacttcaaga
     1201 tccgccacaa catcgaggac ggcggcgtgc agctcgccga ccactaccag cagaacaccc
     1261 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc
     1321 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     1381 ccgggatcac tctcggcatg gacgagctgt acaagtaagc ggccgcactc ctcaggtgca
     1441 ggctgcctat cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca aataccactg
     1501 agatcttttt ccctctgcca aaaattatgg ggacatcatg aagccccttg agcatctgac
     1561 ttctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct
     1621 ctcactcgga aggacatatg ggagggcaaa tcatttaaaa catcagaatg agtatttggt
     1681 ttagagtttg gcaacatatg cccatatgct ggctgccatg aacaaaggtt ggctataaag
     1741 aggtcatcag tatatgaaac agccccctgc tgtccattcc ttattccata gaaaagcctt
     1801 gacttgaggt tagatttttt ttatattttg ttttgtgtta tttttttctt taacatccct
     1861 aaaattttcc ttacatgttt tactagccag atttttcctc ctctcctgac tactcccagt
     1921 catagctgtc cctcttctct tatggagatc cctcgacctg cagcccaagc ttggcgtaat
     1981 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     2041 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     2101 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag cggatccgca
     2161 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     2221 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     2281 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     2341 aggcttttgc aaaaagctaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     2401 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     2461 aaactcatca atgtatctta tcatgtctgg atccgctgca ttaatgaatc ggccaacgcg
     2521 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     2581 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     2641 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     2701 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     2761 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     2821 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     2881 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     2941 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     3001 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     3061 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     3121 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     3181 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     3241 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     3301 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     3361 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     3421 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     3481 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     3541 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     3601 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     3661 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     3721 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     3781 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     3841 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     3901 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     3961 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     4021 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     4081 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     4141 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     4201 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     4261 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     4321 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     4381 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     4441 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgggtcgaca ttgattattg
     4501 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
     4561 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
     4621 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
     4681 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
     4741 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
     4801 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
     4861 accatggtcg aggtgagccc cacgttctgc ttcactctcc ccatctcccc cccctcccca
     4921 cccccaattt tgtatttatt tattttttaa ttattttgtg cagcgatggg ggcggggggg
     4981 gggggggggg cgcgcgccag gcggggcggg gcggggcgag gggcggggcg gggcgaggcg
     5041 gagaggtgcg gcggcagcca atcagagcgg cgcgctccga aagtttcctt ttatggcgag
     5101 gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgcg
     5161 ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact
     5221 gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta
     5281 gcgcttggtt taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc ttgaggggct
     5341 ccgggagggc cctttgtgcg gggggagcgg ctcggggggt gcgtgcgtgt gtgtgtgcgt
     5401 ggggagcgcc gcgtgcggct ccgcgctgcc cggcggctgt gagcgctgcg ggcgcggcgc
     5461 ggggctttgt gcgctccgca gtgtgcgcga ggggagcgcg gccgggggcg gtgccccgcg
     5521 gtgcgggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg tgggggggtg
     5581 agcagggggt gtgggcgcgt cggtcgggct gcaacccccc ctgcaccccc ctccccgagt
     5641 tgctgagcac ggcccggctt cgggtgcggg gctccgtacg gggcgtggcg cggggctcgc
     5701 cgtgccgggc ggggggtggc ggcaggtggg ggtgccgggc ggggcggggc cgcctcgggc
     5761 cggggagggc tcgggggagg ggcgcggcgg cccccggagc gccggcggct gtcgaggcgc
     5821 ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg gacttccttt
     5881 gtcccaaatc tgtgcggagc cgaaatctgg gaggcgccgc cgcaccccct ctagcgggcg
     5941 cggggcgaag cggtgcggcg ccggcaggaa ggaaatgggc ggggagggcc ttcgtgcgtc
     6001 gccgcgccgc cgtccccttc tccctctcca gcctcggggc tgtccgcggg gggacggctg
     6061 ccttcggggg ggacggggca gggcggggtt cggcttctgg cgtgtgaccg gcggctctag
     6121 agcctctgct aaccatgttc atgccttctt ctttttccta cagctcctgg gcaacgtgct
     6181 ggttattgtg ctgtctcatc attttggcaa agaattcctc gagatatcaa acaagtttgt
     6241 acaaaaaagc aggctacacc atggctccaa agaagaagcg taaggtaatg gtgagcaagg
     6301 gcgaggagga caacatggcc atcatcaagg agttcatgcg cttcaaggtg cacatggagg
     6361 gctccgtgaa cggccgcgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg
     6421 gcacccagac cgccaagctg aaggtgacca agggcggccc cctgcccttc gcctgggaca
     6481 tcctgtcccc tcagttcatg tacggctcca aggcctacgt gaagcacccc gccgacatcc
     6541 ccgactactt gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg
     6601 aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc gagttcatct
     6661 acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg cagaagaaga
     6721 ccatgggctg ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg
     6781 agatcaagca gaggctgaag ctgaaggacg gcggccacta cgacgccgag gtcaagacca
     6841 cctacaaggc caagaagccc gtgcagctgc ccggcgccta caacgtcaac atcaagctgg
     6901 acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgagcgc gccgagggcc
     6961 gccactccac cggcggcatg gacgagctgt acaagtaaga cccagcttt
//
