LOCUS       Exported                6882 bp ds-DNA     circular SYN 13-DEC-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6882)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Nov 24, 2020 from SnapGene Viewer 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6882
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     142..158
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     182..219
                     /label=Primer 1
     misc_feature    202..1081
                     /label=SalI arm
     primer_bind     complement(1064..1101)
                     /label=Primer 2
     CDS             1088..1483
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             1493..1549
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1550..1573
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1583..1903
                     /codon_start=1
                     /product="F36V mutant of FK506-binding protein FKBP12"
                     /label=DmrB
                     /note="binds synthetic ligands that do not bind wild-type 
                     FKBP"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             1910..1936
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1937..2650
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     2667..2704
                     /label=Primer 3
     misc_feature    2687..4294
                     /label=KpnI arm
     CDS             2687..2747
                     /codon_start=1
                     /translation="NGDMPHVPITTLAGIASLTD"
     primer_bind     complement(4277..4329)
                     /label=Primer 4
     promoter        complement(4385..4403)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4424..4440)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4448..4464
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(4472..4502)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4517..4538
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4826..5414)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5585..6445)
                     /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(6446..6550)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
       61 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
      121 ggttttccca gtcacgacgt tgtaaaacga cggccagtgc cactagtggc cgcggcggcc
      181 gcagcttgcc tgcaggtcga ccagactctt gtgctgaggg aaagatccta gttccgcaga
      241 ttatcttctg atctgcacac acatcagtca tagcaggcac acaacaatac acgtggacac
      301 taagtgaaca aatagtaaca aattacaaat ggggcagggg aggaggaggg acatatcaag
      361 tgtgtcacaa aggaatatac agcaatttac tgagaaaata ttgggcaata aatgtcataa
      421 attcagttat cctaaggtaa aatggctgtg gaatgaaagt gttcttgaat tgtaggattt
      481 agaagagagg agaataaaag tctcacactg caaaccacaa ggatccaaaa gaatctgaag
      541 agatactctt ggtaggaggg gcaagtccat ttagagagta aaaagggaac tagacaagaa
      601 attgaagcca gattattaag taaaaatttt attgtattta gagtttagaa agagtaccct
      661 gccctagcag tatatacacc agaagttaag aaacaagatt ctggagctag attgtgtcat
      721 tagaattaca gctccagtag ttactttaaa ctcactgtaa ttttctcatt agtaaaatgc
      781 agaaaagagt aatactctgc tgtacctggg ttgttgtgag acctaaatct taatgtatac
      841 aaagcatttt gcagtgttta atgtatggca catgcttgtt aaacacattg aacaaataga
      901 agctaagagt tttgtttcta aatgaataag tatctctctg ctactcatat atttgacaaa
      961 gttctacaaa taattatatg gttttctttg tgtgtgttgc agagtttggg aaatggaaag
     1021 taatgacagc tggcacctga actaaatact ttcatagaca acaccattcc agaacttcag
     1081 ggtcgacatg gccaagcctt tgtctcaaga agaatccacc ctcattgaaa gagcaacggc
     1141 tacaatcaac agcatcccca tctctgaaga ctacagcgtc gccagcgcag ctctctctag
     1201 cgacggccgc atcttcactg gtgtcaatgt atatcatttt actgggggac cttgtgcaga
     1261 actcgtggtg ctgggcactg ctgctgctgc ggcagctggc aacctgactt gtatcgtcgc
     1321 gatcggaaat gagaacaggg gcatcttgag cccctgcgga cggtgccgac aggtgcttct
     1381 cgatctgcat cctgggatca aagccatagt gaaggacagt gatggacagc cgacggcagt
     1441 tgggattcgt gaattgctgc cctctggtta tgtgtgggag ggcggttccg gagccacgaa
     1501 cttctctctg ttaaagcaag caggagacgt ggaagaaaac cccggtcccg attacaagga
     1561 tgacgatgac aagggttccg gcggcgtgca ggtggagact atctccccag gagacgggcg
     1621 caccttcccc aagcgcggcc agacctgcgt ggtgcactac accgggatgc ttgaagatgg
     1681 aaagaaagtt gattcctccc gggacagaaa caagcccttt aagtttatgc taggcaagca
     1741 ggaggtgatc cgaggctggg aagaaggggt tgcccagatg agtgtgggtc agagagccaa
     1801 actgactata tctccagatt atgcctatgg tgccactggg cacccaggca tcatcccacc
     1861 acatgccact ctcgtcttcg atgtggagct tctaaaactg gaagtcgatt atccgtacga
     1921 cgtaccagac tacgcagtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     1981 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     2041 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     2101 gccctggccc accctcgtga ccaccttcgg ctacggcctg cagtgcttcg cccgctaccc
     2161 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     2221 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     2281 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     2341 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
     2401 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
     2461 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     2521 gcccgacaac cactacctga gctaccagtc cgccctgagc aaagacccca acgagaagcg
     2581 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     2641 gctgtacaag ggtggaggtg gatctggtgg cggcggttca ggtaccaatg gggatatgcc
     2701 ccatgtcccc ataactacgc ttgcggggat tgctagtctt acggaccgta agtttgcata
     2761 attttcctaa gttctttact ttgctaatac tcatttttag agacaaaaaa ttacctttaa
     2821 tgtcacatat aacagaaaat atgtatttga ttagtgtggg gggagtagca aaattaatgt
     2881 ttccacctaa tacataaatt taatttcttc atccttctgt gaactcctaa agaaatgata
     2941 gtgtgataat ttttataaag accttaaaaa tcaataagga ctttagaaaa atgacacaaa
     3001 gctttaagag atgttaaaac ttaaataaac atgggttaat aaacaaaact atggtctttt
     3061 ttttaagaaa gcgtttacaa aaacagtacc tgaaagtgtc ctccaagaat gagaaattga
     3121 tcgcccacat tacctaaact gtagtggtcc tgattttttt aatatcaggt tgaattgtta
     3181 gttagaattt gttaacaggt atgagatatg tcatcactat gagagcccac tttactctga
     3241 acactaaggt attaaaaatt attgtcaata cttagaaatt tagttaatag aagttttaag
     3301 gttcatattt tggtagacac ataactattc ttttccttgg gtaatggcac agagtctgcc
     3361 caagtgtttg tccctatgtc atcctccttt tagtatagga atagagataa aggaaaggcc
     3421 agaaaagaaa aaacaaaatg aaaacataaa tacctttttc ttatagcagt atgtaagata
     3481 tggaactttg caataatcta tgaaaaaata acttccagaa ctgttttcaa ctgtgctcag
     3541 aatccctccc tgggttttgt ttttgtgtat tttcagtcga gtgtaagaca cttgtatcac
     3601 tagaaagtag gtggttggat tatcctaaat ctgcagggaa actttgaaac atgtttgttt
     3661 ttgttactcg ttttaaggca aaaacacaac ctgtatggat ggaaaattat atatgctgct
     3721 agacttccct ttatcttttc tgccttttat ctctgatata cctcacattt ttgatgctta
     3781 tcttcctggt aaactctgtg aagtatggta attatactat ggggtcgact agaaattaac
     3841 cccaaaacat gatacctata ttttagttgg gtttcttaac cttttggcaa gtaaacaatt
     3901 cctattaagt ctacttacat taaatagtgg gattgaagcc acaaaatact catttgtttg
     3961 tttgtttgtt tgtttgctgc ttagagtaga atgggagaaa aaagagaact atttaggaat
     4021 ttatgacaag aattttgtat tgttcttcag gaatgttgta ctctgagtat tccactactg
     4081 tgagctattt gtttctatat tcttaagctt ttaaaattgt attgcatatc agtataatgt
     4141 gagtcaggta ttgtttttga cttatatgtc taagagtgct actgatctta tagataagct
     4201 actttctcac cttatgtgtc aaagtcatga taataataat aaagttttat ttagttaatt
     4261 aaaacaattt ttaagcatga atataaatca gctaggtacc gggccccccc tcgaggtcga
     4321 tcgacggtat cgataagctt gatctagagc ggccgccacc gcggtggagc tccagctttt
     4381 gttcccttta gtgagggtta attccgagct tggcgtaatc atggtcatag ctgtttcctg
     4441 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     4501 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     4561 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     4621 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     4681 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     4741 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     4801 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4861 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     4921 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     4981 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     5041 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     5101 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     5161 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     5221 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     5281 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     5341 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     5401 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     5461 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     5521 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     5581 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     5641 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     5701 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     5761 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     5821 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     5881 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     5941 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     6001 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     6061 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     6121 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     6181 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     6241 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     6301 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     6361 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     6421 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     6481 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     6541 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     6601 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
     6661 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     6721 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     6781 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
     6841 aaataccgca cagatgcgta aggagaaaat accgcatcag gc
//
