LOCUS       Exported                7919 bp ds-DNA     circular SYN 03-AUG-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7919)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Jul 7, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     Annotated with pLannotate v1.2.0
FEATURES             Location/Qualifiers
     source          1..7919
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             536..841
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(882..1006)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(1015..1032)
                     /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    1044..1629
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             1630..1689
                     /codon_start=1
                     /label=GAP43 N-term
                     /translation="MLCCMRRTKQVEKNDEDQKI"
     CDS             1690..2409
                     /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    2560..2615
                     /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(2976..2992)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3000..3016)
                     /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(3024..3054)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3069..3090)
                     /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"
     protein_bind    3069..3090
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             complement(3103..3142)
                     /label=lacI (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=3.7"
                     /note="/other=CDS"
     promoter        3149..3344
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3195..3330
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3350..3484
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(3486..3540)
                     /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(3723..4311)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4482..5342)
                     /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(5343..5447)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        5478..5857
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5859..6137
                     /label=chicken beta-actin promoter
     intron          6138..7154
                     /label=chimeric intron
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=99.9"
                     /note="/match_length=100.0"
                     /note="/other=intron"
     intron          6231..7154
                     /label=chicken b-actin intron + rabbit b-globin intron
     misc_feature    7078..7100
                     /label=pCAGGS FWDSeq
     intron          7155..7207
                     /label=rabbit B-globin intron
     protein_bind    7222..7346
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        7371..7401
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     promoter        7425..7454
                     /label=cat promoter (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=29.1"
                     /note="/other=promoter"
     CDS             join(7455..7919,1..216)
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
ORIGIN
        1 tttgatttaa acgtggccaa tatggacaac ttcttcgccc ccgttttcac catgggcaaa
       61 tattatacgc aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt
      121 tgtgatggct tccatgtcgg cagaatgctt aatgaattac aacagtactg cgatgagtgg
      181 caggcggggc gtaatctaga ggatccggct tactaaaagc cagataacag tatgcgtatt
      241 tgcgcgctga tttttgcggt ataagaatat atactgatat gtatacccga agtatgtcaa
      301 aaagaggtat gctatgaagc agcgtattac agtgacagtt gacagcgaca gctatcagtt
      361 gctcaaggca tatatgatgt caatatctcc ggtctggtaa gcacaaccat gcagaatgaa
      421 gcccgtcgtc tgcgtgccga acgctggaaa gcggaaaatc aggaagggat ggctgaggtc
      481 gcccggttta ttgaaatgaa cggctctttt gctgacgaga acaggggctg gtgaaatgca
      541 gtttaaggtt tacacctata aaagagagag ccgttatcgt ctgtttgtgg atgtacagag
      601 tgatattatt gacacgcccg ggcgacggat ggtgatcccc ctggccagtg cacgtctgct
      661 gtcagataaa gtcccccgtg aactttaccc ggtggtgcat atcggggatg aaagctggcg
      721 catgatgacc accgatatgg ccagtgtgcc ggtctccgtt atcggggaag aagtggctga
      781 tctcagccac cgcgaaaatg acatcaaaaa cgccattaac ctgatgttct ggggaatata
      841 aatgtcaggc tcccttatac acagccagtc tgcaggtcga ccatagtgac tggatatgtt
      901 gtgttttaca gtattatgta gtctgttttt tatgcaaaat ctaatttaat atattgatat
      961 ttatatcatt ttacgtttct cgttcagctt tcttgtacaa agtggttcga tatcctatag
     1021 tgtcacctaa ataattccgc ccccccctct ccctcccccc ccctaacgtt actggccgaa
     1081 gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt
     1141 cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg
     1201 gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc
     1261 ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc
     1321 ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa
     1381 aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc
     1441 tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg
     1501 gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac
     1561 gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga tgataatatg
     1621 gccacaacca tgctgtgctg tatgagaaga accaaacagg ttgaaaagaa tgatgaggac
     1681 caaaagatca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     1741 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     1801 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     1861 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
     1921 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     1981 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     2041 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     2101 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     2161 cagaagaacg gcatcaaggc gaacttcaag atccgccaca acatcgagga cggcggcgtg
     2221 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     2281 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     2341 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     2401 tacaagtaag cggccgcact cctcaggtgc aggctgccta tcagaaggtg gtggctggtg
     2461 tggccaatgc cctggctcac aaataccact gagatctttt tccctctgcc aaaaattatg
     2521 gggacatcat gaagcccctt gagcatctga cttctggcta ataaaggaaa tttattttca
     2581 ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg aaggacatat gggagggcaa
     2641 atcatttaaa acatcagaat gagtatttgg tttagagttt ggcaacatat gcccatatgc
     2701 tggctgccat gaacaaaggt tggctataaa gaggtcatca gtatatgaaa cagccccctg
     2761 ctgtccattc cttattccat agaaaagcct tgacttgagg ttagattttt tttatatttt
     2821 gttttgtgtt atttttttct ttaacatccc taaaattttc cttacatgtt ttactagcca
     2881 gatttttcct cctctcctga ctactcccag tcatagctgt ccctcttctc ttatggagat
     2941 ccctcgacct gcagcccaag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     3001 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     3061 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     3121 tcgggaaacc tgtcgtgcca gcggatccgc atctcaatta gtcagcaacc atagtcccgc
     3181 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     3241 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     3301 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagcta acttgtttat
     3361 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     3421 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     3481 gatccgctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     3541 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     3601 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     3661 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     3721 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     3781 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     3841 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     3901 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     3961 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     4021 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     4081 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     4141 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     4201 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     4261 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     4321 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     4381 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     4441 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     4501 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     4561 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     4621 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     4681 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     4741 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     4801 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     4861 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     4921 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     4981 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     5041 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     5101 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     5161 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     5221 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     5281 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     5341 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     5401 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     5461 aaagtgccac ctgggtcgac attgattatt gactagttat taatagtaat caattacggg
     5521 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
     5581 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
     5641 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
     5701 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
     5761 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
     5821 gcagtacatc tacgtattag tcatcgctat taccatggtc gaggtgagcc ccacgttctg
     5881 cttcactctc cccatctccc ccccctcccc acccccaatt ttgtatttat ttatttttta
     5941 attattttgt gcagcgatgg gggcgggggg gggggggggg gcgcgcgcca ggcggggcgg
     6001 ggcggggcga ggggcggggc ggggcgaggc ggagaggtgc ggcggcagcc aatcagagcg
     6061 gcgcgctccg aaagtttcct tttatggcga ggcggcggcg gcggcggccc tataaaaagc
     6121 gaagcgcgcg gcgggcggga gtcgctgcgc gctgccttcg ccccgtgccc cgctccgccg
     6181 ccgcctcgcg ccgcccgccc cggctctgac tgaccgcgtt actcccacag gtgagcgggc
     6241 gggacggccc ttctcctccg ggctgtaatt agcgcttggt ttaatgacgg cttgtttctt
     6301 ttctgtggct gcgtgaaagc cttgaggggc tccgggaggg ccctttgtgc ggggggagcg
     6361 gctcgggggg tgcgtgcgtg tgtgtgtgcg tggggagcgc cgcgtgcggc tccgcgctgc
     6421 ccggcggctg tgagcgctgc gggcgcggcg cggggctttg tgcgctccgc agtgtgcgcg
     6481 aggggagcgc ggccgggggc ggtgccccgc ggtgcggggg gggctgcgag gggaacaaag
     6541 gctgcgtgcg gggtgtgtgc gtgggggggt gagcaggggg tgtgggcgcg tcggtcgggc
     6601 tgcaaccccc cctgcacccc cctccccgag ttgctgagca cggcccggct tcgggtgcgg
     6661 ggctccgtac ggggcgtggc gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg
     6721 gggtgccggg cggggcgggg ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg
     6781 gcccccggag cgccggcggc tgtcgaggcg cggcgagccg cagccattgc cttttatggt
     6841 aatcgtgcga gagggcgcag ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg
     6901 ggaggcgccg ccgcaccccc tctagcgggc gcggggcgaa gcggtgcggc gccggcagga
     6961 aggaaatggg cggggagggc cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc
     7021 agcctcgggg ctgtccgcgg ggggacggct gccttcgggg gggacggggc agggcggggt
     7081 tcggcttctg gcgtgtgacc ggcggctcta gagcctctgc taaccatgtt catgccttct
     7141 tctttttcct acagctcctg ggcaacgtgc tggttattgt gctgtctcat cattttggca
     7201 aagaattcct cgagatatca aacaagtttg tacaaaaaag ctgaacgaga aacgtaaaat
     7261 gatataaata tcaatatatt aaattagatt ttgcataaaa aacagactac ataatactgt
     7321 aaaacacaac atatccagtc actatggcgg ccgcattagg caccccaggc tttacacttt
     7381 atgcttccgg ctcgtataat gtgtggattt tgagttagga tccgtcgaga ttttcaggag
     7441 ctaaggaagc taaaatggag aaaaaaatca ctggatatac caccgttgat atatcccaat
     7501 ggcatcgtaa agaacatttt gaggcatttc agtcagttgc tcaatgtacc tataaccaga
     7561 ccgttcagct ggatattacg gcctttttaa agaccgtaaa gaaaaataag cacaagtttt
     7621 atccggcctt tattcacatt cttgcccgcc tgatgaatgc tcatccggaa ttccgtatgg
     7681 caatgaaaga cggtgagctg gtgatatggg atagtgttca cccttgttac accgttttcc
     7741 atgagcaaac tgaaacgttt tcatcgctct ggagtgaata ccacgacgat ttccggcagt
     7801 ttctacacat atattcgcaa gatgtggcgt gttacggtga aaacctggcc tatttcccta
     7861 aagggtttat tgagaatatg tttttcgtct cagccaatcc ctgggtgagt ttcaccagt
//
