LOCUS       Exported                7896 bp ds-DNA     circular SYN 25-MAY-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7896)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Jul 15, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     Annotated with pLannotate v1.2.0
FEATURES             Location/Qualifiers
     source          1..7896
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      909..1497
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             1680..1734
                     /label=lacI (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=5.1"
                     /note="/other=CDS"
     polyA_signal    1736..1870
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(1876..2071)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1890..2025
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2078..2117
                     /label=lacI (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=3.7"
                     /note="/other=CDS"
     protein_bind    2130..2151
                     /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"
     promoter        2166..2196
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2204..2220
                     /label=lac operator
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=protein_bind"
     primer_bind     2228..2244
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    2605..2660
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     CDS             complement(2811..3509)
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL
                     KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY
                     CDLPSKLGHR"
     CDS             complement(3510..3569)
                     /codon_start=1
                     /label=Gap43 (fragment)
                     /note="pLannotate"
                     /note="/database=swissprot"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=8.8"
                     /note="/other=CDS"
                     /translation="MLCCMRRTKQVEKNDEDQKI"
     misc_feature    3570..4156
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     promoter        4168..4185
                     /label=SP6 promoter (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=94.7"
                     /note="/other=promoter"
     protein_bind    4194..4318
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(4359..4664)
                     /label=ccdB
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=99.3"
                     /note="/match_length=100.0"
                     /note="/other=CDS"
     CDS             complement(4984..5664)
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     promoter        complement(5665..5694)
                     /label=cat promoter (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=29.1"
                     /note="/other=promoter"
     promoter        complement(5718..5748)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(5773..5897)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     intron          complement(5912..5964)
                     /label=rabbit B-globin intron
     intron          complement(5965..6981)
                     /label=chimeric intron
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=99.9"
                     /note="/match_length=100.0"
                     /note="/other=intron"
     intron          complement(5965..6888)
                     /label=chicken b-actin intron + rabbit b-globin intron
     promoter        complement(6982..7257)
                     /label=chicken beta-actin promoter
     enhancer        7259..7638
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7669..7773
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7774..7896,1..738)
                     /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 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
       61 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
      121 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
      181 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
      241 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
      301 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
      361 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
      421 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
      481 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
      541 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
      601 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
      661 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
      721 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
      781 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
      841 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
      901 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
      961 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     1021 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     1081 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     1141 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     1201 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     1261 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     1321 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     1381 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     1441 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     1501 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     1561 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     1621 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     1681 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagcggatcc
     1741 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     1801 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     1861 taaacaagtt agctttttgc aaaagcctag gcctccaaaa aagcctcctc actacttctg
     1921 gaatagctca gaggccgagg cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc
     1981 atggggcgga gaatgggcgg aactgggcgg agttaggggc gggatgggcg gagttagggg
     2041 cgggactatg gttgctgact aattgagatg cggatccgct ggcacgacag gtttcccgac
     2101 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     2161 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     2221 tttcacacag gaaacagcta tgaccatgat tacgccaagc ttgggctgca ggtcgaggga
     2281 tctccataag agaagaggga cagctatgac tgggagtagt caggagagga ggaaaaatct
     2341 ggctagtaaa acatgtaagg aaaattttag ggatgttaaa gaaaaaaata acacaaaaca
     2401 aaatataaaa aaaatctaac ctcaagtcaa ggcttttcta tggaataagg aatggacagc
     2461 agggggctgt ttcatatact gatgacctct ttatagccaa cctttgttca tggcagccag
     2521 catatgggca tatgttgcca aactctaaac caaatactca ttctgatgtt ttaaatgatt
     2581 tgccctccca tatgtccttc cgagtgagag acacaaaaaa ttccaacaca ctattgcaat
     2641 gaaaataaat ttcctttatt agccagaagt cagatgctca aggggcttca tgatgtcccc
     2701 ataatttttg gcagagggaa aaagatctca gtggtatttg tgagccgggg cattggccac
     2761 accagccacc accttctgat aggcagcctg cacctgagga gtgcggccgc tcatctgtgc
     2821 cccagtttgc tagggaggtc gcagtatctg gccacagcca cctcgtgctg ctcgacgtag
     2881 gtctctttgt cggcctcctt gattctttcc agtcttctgt ccacatagta gacgccgggc
     2941 atcttgaggt tcttagcggg tttcttggat ctgtatgtgg tcttcaagtt gcagatcagg
     3001 tggcccccgc ccacgagctt cagggccatg tcggctctgc cttccaggcc gccgtcagcg
     3061 gggtacaggg tctcggtgga ggcctcccag ccgagtgttt tcttctgcat cacagggccg
     3121 ttggatggga agttcacccc tctgatcttg acgttgtaga tgaggcagcc gtcctggagg
     3181 ctggtgtcct gggtagcggt cagcacgccc ccgtcttcgt atgtggtgac tctctcccat
     3241 gtgaagccct cggggaagga ctgcttaaag aagtcgggga tgccctgggt gtggttgatg
     3301 aaggttttgc tgccgtacat gaagctggta gccaggatgt cgaaggcgaa ggggagaggg
     3361 ccgccctcga ccgccttgat tctcatggtc tgggtgccct cgtagggctt gccttcgccc
     3421 tcggatgtgc acttgaagtg gtggttgttc acggtgccct ccatgtacag cttcatgtgc
     3481 atgttctcct taatcagctc gctcaccatg atcttttggt cctcatcatt cttttcaacc
     3541 tgtttggttc ttctcataca gcacagcatg gttgtggcca tattatcatc gtgtttttca
     3601 aaggaaaacc acgtccccgt ggttcggggg gcctagacgt ttttttaacc tcgactaaac
     3661 acatgtaaag catgtgcacc gaggccccag atcagatccc atacaatggg gtaccttctg
     3721 ggcatccttc agccccttgt tgaatacgct tgaggagagc catttgactc tttccacaac
     3781 tatccaactc acaacgtggc actggggttg tgccgccttt gcaggtgtat cttatacacg
     3841 tggcttttgg ccgcagaggc acctgtcgcc aggtgggggg ttccgctgcc tgcaaagggt
     3901 cgctacagac gttgtttgtc ttcaagaagc ttccagagga actgcttcct tcacgacatt
     3961 caacagacct tgcattcctt tggcgagagg ggaaagaccc ctaggaatgc tcgtcaagaa
     4021 gacagggcca ggtttccggg ccctcacatt gccaaaagac ggcaatatgg tggaaaataa
     4081 catatagaca aacgcacacc ggccttattc caagcggctt cggccagtaa cgttaggggg
     4141 gggggaggga gagggggggg cggaattatt taggtgacac tataggatat cgaaccactt
     4201 tgtacaagaa agctgaacga gaaacgtaaa atgatataaa tatcaatata ttaaattaga
     4261 ttttgcataa aaaacagact acataatact gtaaaacaca acatatccag tcactatggt
     4321 cgacctgcag actggctgtg tataagggag cctgacattt atattcccca gaacatcagg
     4381 ttaatggcgt ttttgatgtc attttcgcgg tggctgagat cagccacttc ttccccgata
     4441 acggagaccg gcacactggc catatcggtg gtcatcatgc gccagctttc atccccgata
     4501 tgcaccaccg ggtaaagttc acgggggact ttatctgaca gcagacgtgc actggccagg
     4561 gggatcacca tccgtcgccc gggcgtgtca ataatatcac tctgtacatc cacaaacaga
     4621 cgataacggc tctctctttt ataggtgtaa accttaaact gcatttcacc agcccctgtt
     4681 ctcgtcagca aaagagccgt tcatttcaat aaaccgggcg acctcagcca tcccttcctg
     4741 attttccgct ttccagcgtt cggcacgcag acgacgggct tcattctgca tggttgtgct
     4801 taccagaccg gagatattga catcatatat gccttgagca actgatagct gtcgctgtca
     4861 actgtcactg taatacgctg cttcatagca tacctctttt tgacatactt cgggtataca
     4921 tatcagtata tattcttata ccgcaaaaat cagcgcgcaa atacgcatac tgttatctgg
     4981 cttttagtaa gccggatcct ctagattacg ccccgcctgc cactcatcgc agtactgttg
     5041 taattcatta agcattctgc cgacatggaa gccatcacaa acggcatgat gaacctgaat
     5101 cgccagcggc atcagcacct tgtcgccttg cgtataatat ttgcccatgg tgaaaacggg
     5161 ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa ctggtgaaac tcacccaggg
     5221 attggctgag acgaaaaaca tattctcaat aaacccttta gggaaatagg ccaggttttc
     5281 accgtaacac gccacatctt gcgaatatat gtgtagaaac tgccggaaat cgtcgtggta
     5341 ttcactccag agcgatgaaa acgtttcagt ttgctcatgg aaaacggtgt aacaagggtg
     5401 aacactatcc catatcacca gctcaccgtc tttcattgcc atacggaatt ccggatgagc
     5461 attcatcagg cgggcaagaa tgtgaataaa ggccggataa aacttgtgct tatttttctt
     5521 tacggtcttt aaaaaggccg taatatccag ctgaacggtc tggttatagg tacattgagc
     5581 aactgactga aatgcctcaa aatgttcttt acgatgccat tgggatatat caacggtggt
     5641 atatccagtg atttttttct ccattttagc ttccttagct cctgaaaatc tcgacggatc
     5701 ctaactcaaa atccacacat tatacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     5761 taatgcggcc gccatagtga ctggatatgt tgtgttttac agtattatgt agtctgtttt
     5821 ttatgcaaaa tctaatttaa tatattgata tttatatcat tttacgtttc tcgttcagct
     5881 tttttgtaca aacttgtttg atatctcgag gaattctttg ccaaaatgat gagacagcac
     5941 aataaccagc acgttgccca ggagctgtag gaaaaagaag aaggcatgaa catggttagc
     6001 agaggctcta gagccgccgg tcacacgcca gaagccgaac cccgccctgc cccgtccccc
     6061 ccgaaggcag ccgtcccccc gcggacagcc ccgaggctgg agagggagaa ggggacggcg
     6121 gcgcggcgac gcacgaaggc cctccccgcc catttccttc ctgccggcgc cgcaccgctt
     6181 cgccccgcgc ccgctagagg gggtgcggcg gcgcctccca gatttcggct ccgcacagat
     6241 ttgggacaaa ggaagtccct gcgccctctc gcacgattac cataaaaggc aatggctgcg
     6301 gctcgccgcg cctcgacagc cgccggcgct ccgggggccg ccgcgcccct cccccgagcc
     6361 ctccccggcc cgaggcggcc ccgccccgcc cggcaccccc acctgccgcc accccccgcc
     6421 cggcacggcg agccccgcgc cacgccccgt acggagcccc gcacccgaag ccgggccgtg
     6481 ctcagcaact cggggagggg ggtgcagggg gggttgcagc ccgaccgacg cgcccacacc
     6541 ccctgctcac ccccccacgc acacaccccg cacgcagcct ttgttcccct cgcagccccc
     6601 cccgcaccgc ggggcaccgc ccccggccgc gctcccctcg cgcacactgc ggagcgcaca
     6661 aagccccgcg ccgcgcccgc agcgctcaca gccgccgggc agcgcggagc cgcacgcggc
     6721 gctccccacg cacacacaca cgcacgcacc ccccgagccg ctccccccgc acaaagggcc
     6781 ctcccggagc ccctcaaggc tttcacgcag ccacagaaaa gaaacaagcc gtcattaaac
     6841 caagcgctaa ttacagcccg gaggagaagg gccgtcccgc ccgctcacct gtgggagtaa
     6901 cgcggtcagt cagagccggg gcgggcggcg cgaggcggcg gcggagcggg gcacggggcg
     6961 aaggcagcgc gcagcgactc ccgcccgccg cgcgcttcgc tttttatagg gccgccgccg
     7021 ccgccgcctc gccataaaag gaaactttcg gagcgcgccg ctctgattgg ctgccgccgc
     7081 acctctccgc ctcgccccgc cccgcccctc gccccgcccc gccccgcctg gcgcgcgccc
     7141 cccccccccc ccgcccccat cgctgcacaa aataattaaa aaataaataa atacaaaatt
     7201 gggggtgggg agggggggag atggggagag tgaagcagaa cgtggggctc acctcgacca
     7261 tggtaatagc gatgactaat acgtagatgt actgccaagt aggaaagtcc cataaggtca
     7321 tgtactgggc ataatgccag gcgggccatt taccgtcatt gacgtcaata gggggcgtac
     7381 ttggcatatg atacacttga tgtactgcca agtgggcagt ttaccgtaaa tactccaccc
     7441 attgacgtca atggaaagtc cctattggcg ttactatggg aacatacgtc attattgacg
     7501 tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc atttaccgta agttatgtaa
     7561 cgcggaactc catatatggg ctatgaacta atgaccccgt aattgattac tattaataac
     7621 tagtcaataa tcaatgtcga cccaggtggc acttttcggg gaaatgtgcg cggaacccct
     7681 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     7741 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     7801 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     7861 aaagtaaaag atgctgaaga tcagttgggt gcacga
//
