LOCUS       CLYBL-Bxb1-LP-v2        8531 bp DNA     circular SYN 10-NOV-2022
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 8531)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Nov 10, 2022 from SnapGene 6.1.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8531
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     misc_feature    147..979
                     /label=CLYBL LA
                     /note="color: #00ffff"
     misc_feature    1014..1033
                     /label=MCS
                     /note="MCS"
                     /note="color: #0080ff"
     protein_bind    1034..1066
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
                     /note="color: #31849b; direction: RIGHT"
     promoter        1122..1621
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /note="color: #ffffff; direction: RIGHT"
     CDS             1636..2352
                     /codon_start=1
                     /product="enhanced blue variant of GFP (Ai et al., 2007)"
                     /label=EBFP2
                     /note="mammalian codon-optimized"
                     /note="This feature has 3 segments:
                        1: 1636 .. 1638 / #3366ff
                        2: 1639 .. 1641 / #3366ff / 1a
                        3: 1642 .. 2352 / #3366ff"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
                     VNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL
                     EFRTAAGITLGMDELYK"
     CDS             2360..2362
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /note="color: #05fd14"
                     /translation=""
     polyA_signal    2405..2526
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     protein_bind    2545..2592
                     /label=attP
                     /bound_moiety="bxb1 integrase"
                     /note="color: #31849b; direction: RIGHT"
     CDS             2598..2966
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /note="color: #ccffcc"
                     /translation="AKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDDV
                     TLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGRE
                     FALRDPAGNCVHFVAEEQD"
     polyA_signal    2975..3497
                     /label=polyA
                     /note="Rabbit bGlobulin polyAd signal"
                     /note="color: #ff0080"
     protein_bind    3559..3591
                     /label=lox251
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites."
                     /note="color: #31849b; direction: RIGHT"
     misc_feature    3619..4415
                     /label=CLYBL RA
                     /note="color: #00ffff"
     rep_origin      4563..5273
                     /label=oriV
                     /note="incP origin of replication"
                     /note="color: #ffff00"
     promoter        5294..5398
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             5399..6193
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /note="color: #ccffcc"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      6364..6952
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     rep_origin      6952
                     /label=ORI
                     /note="ORI"
                     /note="RNaseH cleavage point"
                     /note="color: #3366cc"
     CDS             7288..8436
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /note="color: #993366"
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
ORIGIN
        1 agtggtgatt ttgtgccgag ctgccggtcg gggagctgtt ggctggctgg tggcaggata
       61 tattgtggtg taaacaaatt gacgcttaga caacttaata acacattgcg gacgttttta
      121 atgtactggg gttgaacact ctgtgccagc aggagaatca tgccaatggg ccaatataca
      181 ttctgaccca cagtttcata ataaaataaa atggttgtgg ttgtaagcca ctatgtttca
      241 gagtggtttg ttacacagca ataaataact aatatagtag gcataccatc aagtccaaag
      301 taggtagaga agaatgtaaa tagcagagca aaacagcatg actggtggct gggaggctta
      361 aaactgggac aggatcagag tcatgaaaga agtcaaagaa atggttcaga agtaaggctg
      421 agactgactt acaaaagctg aaagtccctt taagttggtg tttggtgcat tggcaggggc
      481 aggtatggtg acttaaaaga gccatgctca acaagatcaa gcacaacaca atcacgggtc
      541 accccagcag accttagcga gtctagccat ttctttggtg gtggtcacag tcatgcttca
      601 gcccagtttc cacttggaca aatggtacat attttcaatg agatgaaaat taagatacaa
      661 tccatgtgct cagagagtga tcacagctct gactaaacac tgtgccccaa agtgttgagg
      721 aattgggaaa acctagctga gttagtggtc tcttttctgt tacaataaag ctcataatga
      781 aaattagcct tctttgttct tccccaagtc tttctttcta gacgaaacta ctttcaactg
      841 ttttaacttc cttactgtta acttccatat ttctagatag tatggtcaga ctgttatttc
      901 ttgactttta aatgtaagat attatctact gacttccttc tatgtaagat gaggattgag
      961 ctctcttacc cttctcccaa ctaagtcacg acgttgtaaa acgacggcca gtcgctagcg
     1021 agctcggcgc gccataactt cgtatagcat acattatacg aagttagaat tcgccttctg
     1081 caggagcgta cagaacccag ggccctggca cccgtgctac cgggtagggg aggcgctttt
     1141 cccaaggcag tctggagcat gcgctttagc agccccgctg ggcacttggc gctacacaag
     1201 tggcctctgg cctcgcacac attccacatc caccggtagg cgccaaccgg ctccgttctt
     1261 tggtggcccc ttcgcgccac cttccactcc tcccctagtc aggaagttcc cccccgcccc
     1321 gcagctcgcg tcgtgcagga cgtgacaaat ggaagtagca cgtctcacta gtctcgtgca
     1381 gatggacagc accgctgagc aatggaagcg ggtaggcctt tggggcagcg gccaatagca
     1441 gctttgctcc ttcgctttct gggctcagag gctgggaagg ggtgggtccg ggggcgggct
     1501 caggggcggg ctcaggggcg gggcgggcgc ccgaaggtcc tccggaggcc cggcattctg
     1561 cacgcttcaa aagcgcacgt ctgccgcgct gttctcctct tcctcatctc cgggcctttc
     1621 gacctgcagg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     1681 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgagggg cgagggcgag
     1741 ggcgatgcca ccaacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     1801 gtgccctggc ccaccctcgt gaccaccctg agccacggcg tgcagtgctt cgcccgctac
     1861 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     1921 gagcgcacca tcttcttcaa ggacgacggc acctacaaga cccgcgccga ggtgaagttc
     1981 gagggcgaca ccctagtgaa ccgcatcgag ctgaagggcg tcgacttcaa ggaggacggc
     2041 aacatcctgg ggcacaagct ggagtacaac ttcaacagcc acaacatcta tatcatggcc
     2101 gtcaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacgt ggaggacggc
     2161 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     2221 ctgcccgaca gccactacct gagcacccag tccgtgctga gcaaagaccc caacgagaag
     2281 cgcgatcaca tggtcctgct ggagttccgc accgccgccg ggatcactct cggcatggac
     2341 gagctgtaca agtaatcgat aactgcagcg cggggatctc atgctggagt tcttcgccca
     2401 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     2461 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     2521 atcttatcat gtctagatat acgtggtttg tctggtcaac caccgcggtc tcagtggtgt
     2581 acggtacaaa cccatatgcc aagttgacca gtgccgttcc ggtgctcacc gcgcgcgacg
     2641 tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac ttcgtggagg
     2701 acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg gtccaggacc
     2761 aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg cggcctggac gagctgtacg
     2821 ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc ctccgggccg gccatgaccg
     2881 agatcggcga gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc ggcaactgcg
     2941 tgcacttcgt ggccgaggag caggactgac attcactcct caggtgcagg ctgcctatca
     3001 gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa taccactgag atctttttcc
     3061 ctctgccaaa aattatgggg acatcatgaa gccccttgag catctgactt ctggctaata
     3121 aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct cactcggaag
     3181 gacatatggg agggcaaatc atttaaaaca tcagaatgag tatttggttt agagtttggc
     3241 aacatatgcc catatgctgg ctgccatgaa caaaggtggc tataaagagg tcatcagtat
     3301 atgaaacagc cccctgctgt ccattcctta ttccatagaa aagccttgac ttgaggttag
     3361 atttttttta tattttgttt tgtgttattt ttttctttaa catccctaaa attttcctta
     3421 catgttttac tagccagatt tttcctcctc tcctgactac tcccagtcat agctgtccct
     3481 cttctcttat gaagatcgga tcgctactag aggatgcaca tgtgaccgag ggaggagctg
     3541 caagaactct tcctcacgat aacttcgtat aggagacttt atacgaagtt aagcgcttcg
     3601 tgagaccacg aagtagtcct cctcatcctt ccaacataaa tatattttgg gattatatca
     3661 acattcaatg ttacttaaag tgaccttgta aatattttca caactgagcc atgtttgatt
     3721 tgtatactta tgtttacttt actgtttttc ctgaagttaa taattgcctt gaatttattt
     3781 atttctttaa aaatgtttca ttactcagga ctgtagttta cattacgatt ctttgtgtta
     3841 tacagttgat gggtttcttt tctttcttaa tttctttaaa aaatagagat ggggtcttac
     3901 tatattaccc aggctggtct tgaagtcctg ggctcaagtg atcttcctgt ctcagcctac
     3961 caagtagctg agactatagg tgcaaaaaag ccactatacc tggctagttt acaggtttta
     4021 acaaatgcat tatgccacgt atccattatt acaggatcac acaagatatt ttcattaccc
     4081 tgaaagcatc cctgtgttcc accaattcat cctgcctcca tgagccgctg gcaaccactg
     4141 atctctatag ttttgccttt tctaaaatgt catataattg gaatcataca gtctgtagca
     4201 ttttcagact agcttttaaa atttggcaat atgcatttaa ggttcctcct taaatgtgaa
     4261 gggcatagcc aatgtggctt gatagctcat ttctttttat tggtgaatat ttcattgtct
     4321 ggatgttcca cagtttgttt atccattcac ctattcaatt tgcttttttt ctgtgtatct
     4381 atcactaatt caatactgga ctctccaaca gagccgggac acgaagtgat ccgtttaaac
     4441 tatcagtgtt tgacaggata tattggcggg taaacctaag agaaaagagc gtttattaga
     4501 ataatcggat atttaaaagg gcgtgaaaag gtttatccgt tcgtccattt gtatgtgcca
     4561 gccgcctttg cgacgctcac cgggctggtt gccctcgccg ctgggctggc ggccgtctat
     4621 ggccctgcaa acgcgccaga aacgccgtcg aagccgtgtg cgagacaccg cggccgccgg
     4681 cgttgtggat acctcgcgga aaacttggcc ctcactgaca gatgaggggc ggacgttgac
     4741 acttgagggg ccgactcacc cggcgcggcg ttgacagatg aggggcaggc tcgatttcgg
     4801 ccggcgacgt ggagctggcc agcctcgcaa atcggcgaaa acgcctgatt ttacgcgagt
     4861 ttcccacaga tgatgtggac aagcctgggg ataagtgccc tgcggtattg acacttgagg
     4921 ggcgcgacta ctgacagatg aggggcgcga tccttgacac ttgaggggca gagtgctgac
     4981 agatgagggg cgcacctatt gacatttgag gggctgtcca caggcagaaa atccagcatt
     5041 tgcaagggtt tccgcccgtt tttcggccac cgctaacctg tcttttaacc tgcttttaaa
     5101 ccaatattta taaaccttgt ttttaaccag ggctgcgccc tgtgcgcgtg accgcgcacg
     5161 ccgaaggggg gtgccccccc ttctcgaacc ctcccggccc gctaacgcgg gcctcccatc
     5221 cccccagggg ctgcgcccct cggccgcgaa cggcctcacc ccaaaaatgg cagcgctggc
     5281 caattcccga gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     5341 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     5401 ggctaaaatg agaatatcac cggaattgaa aaaactgatc gaaaaatacc gctgcgtaaa
     5461 agatacggaa ggaatgtctc ctgctaaggt atataagctg gtgggagaaa atgaaaacct
     5521 atatttaaaa atgacggaca gccggtataa agggaccacc tatgatgtgg aacgggaaaa
     5581 ggacatgatg ctatggctgg aaggaaagct gcctgttcca aaggtcctgc actttgaacg
     5641 gcatgatggc tggagcaatc tgctcatgag tgaggccgat ggcgtccttt gctcggaaga
     5701 gtatgaagat gaacaaagcc ctgaaaagat tatcgagctg tatgcggagt gcatcaggct
     5761 ctttcactcc atcgacatat cggattgtcc ctatacgaat agcttagaca gccgcttagc
     5821 cgaattggat tacttactga ataacgatct ggccgatgtg gattgcgaaa actgggaaga
     5881 ggacactcca tttaaagatc cgcgcgagct gtatgatttt ttaaagacgg aaaagcccga
     5941 agaggaactt gtcttttccc acggcgacct gggagacagc aacatctttg tgaaagatgg
     6001 caaagtaagt ggctttattg atcttgggag aagcggcagg gcggacaagt ggtatgacat
     6061 tgccttctgc gtccggtcgc tcagggagga tatcggggaa gaacagtatg tcgagctatt
     6121 ttttgactta ctggggatca agcctgattg ggagaaaata aaatattata ttttactgga
     6181 tgaattgttt tagctgtcag accaagttta ctcatatata ctttagattg atttaaaact
     6241 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     6301 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     6361 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     6421 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     6481 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     6541 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     6601 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     6661 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     6721 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     6781 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     6841 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     6901 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     6961 caacgcggcc tttttacggt tcctggcaga tcctagatgt ggcgcaacga tgccggcgac
     7021 aagcaggagc gcaccgactt cttccgcatc aagtgttttg gctctcaggc cgaggcccac
     7081 ggcaagtatt tgggcaaggg gtcgctggta ttcgtgcagg gcaagattcg gaataccaag
     7141 tacgagaagg acggccagac ggtctacggg accgacttca ttgccgataa ggtggattat
     7201 ctggacacca aggcaccagg cgggtcaaat caggaataag ggcacattgc cccggcgtga
     7261 gtcggggcaa tcccgcaagg agggtgaatg aatcggacgt ttgaccggaa ggcatacagg
     7321 caagaactga tcgacgcggg gttttccgcc gaggatgccg aaaccatcgc aagccgcacc
     7381 gtcatgcgtg cgccccgcga aaccttccag tccgtcggct cgatggtcca gcaagctacg
     7441 gccaagatcg agcgcgacag cgtgcaactg gctccccctg ccctgcccgc gccatcggcc
     7501 gccgtggagc gttcgcgtcg tcttgaacag gaggcggcag gtttggcgaa gtcgatgacc
     7561 atcgacacgc gaggaactat gacgaccaag aagcgaaaaa ccgccggcga ggacctggca
     7621 aaacaggtca gcgaggccaa gcaggccgcg ttgctgaaac acacgaagca gcagatcaag
     7681 gaaatgcagc tttccttgtt cgatattgcg ccgtggccgg acacgatgcg agcgatgcca
     7741 aacgacacgg cccgctctgc cctgttcacc acgcgcaaca agaaaatccc gcgcgaggcg
     7801 ctgcaaaaca aggtcatttt ccacgtcaac aaggacgtga agatcaccta caccggcgtc
     7861 gagctgcggg ccgacgatga cgaactggtg tggcagcagg tgttggagta cgcgaagcgc
     7921 acccctatcg gcgagccgat caccttcacg ttctacgagc tttgccagga cctgggctgg
     7981 tcgatcaatg gccggtatta cacgaaggcc gaggaatgcc tgtcgcgcct acaggcgacg
     8041 gcgatgggct tcacgtccga ccgcgttggg cacctggaat cggtgtcgct gctgcaccgc
     8101 ttccgcgtcc tggaccgtgg caagaaaacg tcccgttgcc aggtcctgat cgacgaggaa
     8161 atcgtcgtgc tgtttgctgg cgaccactac acgaaattca tatgggagaa gtaccgcaag
     8221 ctgtcgccga cggcccgacg gatgttcgac tatttcagct cgcaccggga gccgtacccg
     8281 ctcaagctgg aaaccttccg cctcatgtgc ggatcggatt ccacccgcgt gaagaagtgg
     8341 cgcgagcagg tcggcgaagc ctgcgaagag ttgcgaggca gcggcctggt ggaacacgcc
     8401 tgggtcaatg atgacctggt gcattgcaaa cgctagggcc ttgtggggtc agttccggct
     8461 gggggttcag cagcccctgc tcggatctgt tggaccggac agtagtcatg gttgatgggc
     8521 tgcctgtatc g
//
