LOCUS       Exported                9108 bp ds-DNA     circular SYN 20-JUN-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9108)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Jul 10, 2019 from SnapGene Viewer 4.3.10
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9108
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    107..128
                     /label=CAP binding site
     promoter        143..173
                     /label=lac promoter
     protein_bind    181..197
                     /label=lac operator
     primer_bind     205..221
                     /label=M13 rev
     promoter        239..257
                     /label=SP6 promoter
     misc_feature    294..1834
                     /label=actB5'arm
     CDS             join(294..406,849..1287,1381..1562,1673..1816)
                     /codon_start=1
                     /label=ACTB
                     /translation="IWHHTFYNELRVAPEEHPVLLTEAPLNPKANREKMTQIMFETFNT
                     PAMYVAIQAVLSLYASGRTTGIVMDSGDGVTHTVPIYEGYALPHAILRLDLAGRDLTDY
                     LMKILTERGYSFTTTAEREIVRDIKEKLCYVALDFEQEMATAASSSSLEKSYELPDGQV
                     ITIGNERFRCPEALFQPSFLGMESCGIHETTFNSIMKCDVDIRKDLYANTVLSGGTTMY
                     PGIADRMQKEITALAPSTMKIKIIAPPERKYSVWIGGSILASLSTFQQMWISKQEYDES
                     GPSIVHRKCF"
     misc_feature    294..406
                     /label=Exon2
     misc_feature    849..1287
                     /label=Exon3
     misc_feature    1381..1562
                     /label=Exon4
     misc_feature    1673..1816
                     /label=last Exon
     protein_bind    1848..1881
                     /label=loxP
     misc_feature    join(1906..2388,2390..2483)
                     /label=IRES2
     misc_feature    1910..2483
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             2500..3216
                     /codon_start=1
                     /label=EGFPdelSTOP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             3241..3306
                     /codon_start=1
                     /product="2A peptide from foot-and-mouth disease virus 
                     polyprotein"
                     /label=F2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="VKQTLNFDLLKLAGDVESNPGP"
     CDS             3313..4107
                     /codon_start=1
                     /label=NeoR/KanR
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     protein_bind    4125..4158
                     /label=lox2272
     misc_feature    4181..5977
                     /label=actB3'arm
     misc_feature    4216..4221
                     /label=polyAsignal
     promoter        complement(5996..6014)
                     /label=T7 promoter
     primer_bind     complement(6021..6037)
                     /label=M13 fwd
     CDS             6175..6477
                     /codon_start=1
                     /label=ccdB
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     promoter        6688..6737
                     /label=NEOKAN promoter
     CDS             6826..7620
                     /codon_start=1
                     /label=NeoR/KanR
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        7754..7782
                     /label=AmpR promoter
     CDS             7827..8201
                     /codon_start=1
                     /label=BleoR
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     rep_origin      8339..8927
                     /direction=RIGHT
                     /label=ori
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctat
      241 ttaggtgaca ctatagaata ctcaagctat gcatcaagct tggtaccgag ctcagatctg
      301 gcaccacact ttttacaacg agctgcgtgt ggctcctgag gagcaccccg tgctgcttac
      361 tgaggccccc ctgaacccca aggccaaccg tgagaagatg acccaggtga gtgatctgcc
      421 acctcccctg gtggccgcct ccctccttcc tggcctcccg gagctgcgcc ctttctcact
      481 ggttctctct tctgccgttc tccgtaggac tctcttctct gacctgagtc tcctttggaa
      541 ctctgcaggt tctatttgct ttttcccaga tgaggtcttt ctggtgtttg cctttctgac
      601 taggtgtcta aaacagacag tgctgtgggt gtaggtacta acactggctc gtgtgacaag
      661 cccatgaggc tggtgtaaag cagtcttgga gtgtgtatac agtaggcgca cagtaggtct
      721 gaacagaatc cccatcccaa gaccccagca caactagcgg tgttcttgca cttactgcat
      781 gtcccctgtc tgctcttggc tgtccctagt gacttcctca gtgtgacacg gtgcatctct
      841 ttctacagat catgtttgag accttcaaca ccccagccat gtatgtggcc atccaggctg
      901 tgctttcctt gtacgcctct ggccgtacca ctggcatcgt gatggactcc ggtgacgggg
      961 tcacccacac tgtgcccatc tatgaggggt atgccctccc ccacgccatc ctacgtctgg
     1021 acctggctgg ccgggacctg actgactacc tcatgaagat cctcaccgag cgcggctaca
     1081 gcttcaccac cacagctgag cgggaaatcg tgcgtgacat caaggagaag ctgtgctacg
     1141 tcgccctgga cttcgagcag gagatggcca cggcggcctc cagctcctcc ctggagaaga
     1201 gctatgagct gcccgatggt caggtcatca ccatcggcaa cgagcggttc cgctgccctg
     1261 aggcactctt ccagccttct ttcctgggta agtggagacc actgcctagc cctgcctgac
     1321 atgaggttac cttggggctg tgctgtggaa gcttaagtct gggtctcatt tccccctcag
     1381 gcatggaatc ttgtggcatc cacgaaacta cctttaactc catcatgaaa tgtgacgtgg
     1441 acatccgcaa agacctgtat gccaacacag tgctgtctgg tggtaccacc atgtaccctg
     1501 gcatcgctga caggatgcag aaggagatca ccgccctggc acccagcaca atgaagatca
     1561 aggtgggtgt ccttccagcc tgagctgacc tgggcagggg gcatggggcc ctgtggtttg
     1621 tggggagctg tcatatcaag ggtcctcact gcctgttccc tccactcctc agatcattgc
     1681 tcctcccgag cgcaagtact cagtgtggat tggtggctcc atcctggcct cgctgtccac
     1741 cttccagcag atgtggatca gcaagcagga gtatgatgag tccggcccct ccatcgtcca
     1801 ccgcaaatgc ttctaggcgg actgtgactt agtgaattcg cccttggata acttcgtata
     1861 gcatacatta tacgaagtta tagatcttcc cttttttttc cacagtccgc ccctctccct
     1921 cccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct
     1981 atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc
     2041 ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc
     2101 tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg
     2161 tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa
     2221 agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt
     2281 ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg
     2341 atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtac acatgcttta
     2401 catgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg gacgtggttt
     2461 tcctttgaaa aacacgatga taaggatctg ctagccacca tggtgagcaa gggcgaggag
     2521 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     2581 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     2641 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
     2701 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     2761 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     2821 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     2881 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     2941 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
     3001 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     3061 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc
     3121 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     3181 gccgggatca ctctcggcat ggacgagctg tacaagaggc ctcgcgctaa gcgggcaccg
     3241 gtgaaacaga ctttgaattt tgaccttctc aagttggcgg gagacgtgga gtccaaccca
     3301 gggccggata tcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     3361 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     3421 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     3481 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     3541 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     3601 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     3661 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     3721 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     3781 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     3841 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     3901 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     3961 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     4021 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     4081 tatcgccttc ttgacgagtt cttctgagcg gccggccgct cgacataact tcgtatagga
     4141 tactttatac gaagttatga tatccatcac actggcggcc gcgggcttac ctgtacactg
     4201 acttgagacc agttgaataa agtgcacacc ttaaaaatga ggccaagcat gactttgtgt
     4261 cgggctgggg tgtgggcagc aggtgaaccc aggggaaggg tggggcagtg ggggccagcc
     4321 tgtccctact gagtggaggt gggctgaggt tctctctcct tgacattgag cagccttaga
     4381 atgtgaggga agtttaagat tcaggcctct gctcattggg gagttcctgg cttggccctg
     4441 ccacctgtcc ctaggtaccc cagtttctct gggttcatcc agaaggtcta gatgctcagg
     4501 aagtgctggt tggcacctgt gtttcttagg agtggagatc ccttagggtc actccgggca
     4561 gggggttctt gagtgggagt gtagtgggtg ttcctagctg ccatgccttt gtcctgctca
     4621 ctgtattgga gtggagtcgt ccataggctg agagaagcca tgactcctgc ttctctaccc
     4681 agacagggaa atctgggatc acagtcgaat tgtggtgggg gatccatggg gccatgtggt
     4741 ggcaggcagc tctggttagc tctggactga gctggattct gccttgtgac gagtaggcgc
     4801 catcctgtgg cctctcaggg cccagtttgt gacccatgct gcagagggag atggccagac
     4861 tcctggacag cagggcatgc ctggcattgc aggtcagatt cttttttttt tgagatgggg
     4921 tctggctttg ttgctcaggc tttagttcag tggtacaacc acagctcact gcagcctcag
     4981 ccttctgggc aagcagttct gcctcagcct cccaaatagc tggaactaca caggcacacc
     5041 cagctagttg tgggtcagat tctcacttgt gcagagctga gtcacttttg aaagaaacaa
     5101 aacctggctt cccttccgct gctagaagct ttgattgata gtagcggaga atgaaggaag
     5161 gagacgaggc tcccttgcgg ggataccctt taccaaaaac accacctggt gtatcacagt
     5221 tgttctggaa atgatctggt ccccaggagg cctggactgc ctctcacttg tgcacagtgg
     5281 ctgctggcag agaggaaggt tgtccctttc tgtgttatct tggaggttcc ccgaagggtc
     5341 tgccaagagc agttctagct cttttggtct ttactgagtg accactagat ggtgggcctg
     5401 aagctactga gatgacctgg acaagaaccc tgccttggag gagctgtcaa tctggaaatg
     5461 aggacttcat tcatttgttc attccttgat ccatttctcc attgaatcac tatttgtgct
     5521 atgtactgtg ggtatggagg agaaccaaac agatgtgttc ctgtcctcac caggcaccca
     5581 atttaaaggg gaagacatca taaacacaaa ggtacacaaa tgagtgtcca atgacacggt
     5641 gtcatcatgg gaagaaaaag cagaggcttt gatgaacacg atcattaggt gatattcatt
     5701 atgttgattg tggtggtcta gacataaatt aacacttatc taggctgggc gtggtggctc
     5761 atgcctgtaa tcccagcact ttgggaggct gaggtgggtg gatcaagaga tcgagaccat
     5821 cctggccaac atagtgaaac cccatcttta ctaaaaatac aaaaattagc tgggcgtggt
     5881 ggtgcacact tgggaggctg aggtaggaga actgcttgaa ctggggaggc gaacgttgca
     5941 gtgagctaag attgtgctac tgtactccag cctggtgcta gagggcccaa ttcgccctat
     6001 agtgagtcgt attacaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     6061 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     6121 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctatacgt acggcagttt
     6181 aaggtttaca cctataaaag agagagccgt tatcgtctgt ttgtggatgt acagagtgat
     6241 attattgaca cgccggggcg acggatggtg atccccctgg ccagtgcacg tctgctgtca
     6301 gataaagtct cccgtgaact ttacccggtg gtgcatatcg gggatgaaag ctggcgcatg
     6361 atgaccaccg atatggccag tgtgccggtc tccgttatcg gggaagaagt ggctgatctc
     6421 agccaccgcg aaaatgacat caaaaacgcc attaacctga tgttctgggg aatataaatg
     6481 tcaggcatga gattatcaaa aaggatcttc acctagatcc ttttcacgta gaaagccagt
     6541 ccgcagaaac ggtgctgacc ccggatgaat gtcagctact gggctatctg gacaagggaa
     6601 aacgcaagcg caaagagaaa gcaggtagct tgcagtgggc ttacatggcg atagctagac
     6661 tgggcggttt tatggacagc aagcgaaccg gaattgccag ctggggcgcc ctctggtaag
     6721 gttgggaagc cctgcaaagt aaactggatg gctttctcgc cgccaaggat ctgatggcgc
     6781 aggggatcaa gctctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     6841 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     6901 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     6961 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     7021 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     7081 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     7141 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     7201 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     7261 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     7321 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
     7381 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     7441 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     7501 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     7561 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     7621 attattaacg cttacaattt cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     7681 tcacaccgca tacaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     7741 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     7801 aataatagca cgtgaggagg gccaccatgg ccaagttgac cagtgccgtt ccggtgctca
     7861 ccgcgcgcga cgtcgccgga gcggtcgagt tctggaccga ccggctcggg ttctcccggg
     7921 acttcgtgga ggacgacttc gccggtgtgg tccgggacga cgtgaccctg ttcatcagcg
     7981 cggtccagga ccaggtggtg ccggacaaca ccctggcctg ggtgtgggtg cgcggcctgg
     8041 acgagctgta cgccgagtgg tcggaggtcg tgtccacgaa cttccgggac gcctccgggc
     8101 cggccatgac cgagatcggc gagcagccgt gggggcggga gttcgccctg cgcgacccgg
     8161 ccggcaactg cgtgcacttc gtggccgagg agcaggactg acacgtgcta aaacttcatt
     8221 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     8281 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     8341 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     8401 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     8461 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     8521 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     8581 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     8641 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     8701 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     8761 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     8821 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     8881 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     8941 cggccttttt acggttcctg ggcttttgct ggccttttgc tcacatgttc tttcctgcgt
     9001 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     9061 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaag
//
