LOCUS       Exported                6862 bp ds-DNA     circular SYN 16-AUG-2019
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6862)
  AUTHORS   Bill Lagor
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Aug 16, 2019 from SnapGene Viewer 4.3.10
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6862
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          587..4099
                     /note="color: #ffffff"
     misc_feature    1..185
                     /label=ITR
                     /note="color: #c040ff; direction: RIGHT"
     misc_feature    157
                     /label=Beginning of gblock
                     /note="color: #339966"
     promoter        193..539
                     /label=CBh
                     /note="color: #a6acb3; direction: RIGHT"
     enhancer        193..318
                     /label=Partial CMV enhancer
                     /note="color: #000080; direction: RIGHT"
     promoter        319..539
                     /label=CB promoter
                     /note="color: #66cc99; direction: RIGHT"
     primer_bind     324..344
                     /label=CBh_F
                     /note="color: black; sequence: ttctgcttcactctccccatc; 
                     added: 2019-08-14"
     TATA_signal     515..522
                     /label=TATA
                     /note="color: #0000ff"
     misc_feature    587..592
                     /label=Kozak
                     /note="/__SeqFeature__=True"
                     /note="/__level__=0"
                     /note="/application_notes=Facilitates translation 
                     initiation of ATG start codon downstream of the Kozak 
                     sequence."
                     /note="/full_name=Kozak"
                     /note="/name=Kozak"
                     /note="/official_designation=Kozak"
                     /note="/uuid=5f18b25a1234443494a618363f09f052"
                     /note="/vntifkey=21"
                     /note="color: #800c0c; direction: RIGHT"
     misc_feature    join(593..3436,3446..4099)
                     /label=CreLite
                     /label=SY001
                     /note="This forward directional feature has 5 segments:
                        1:  593 ..  595 / #c0c0c0 / ATG
                        2:  596 ..  616 / #c0c0c0 / NLS
                        3:  617 .. 3436 / #00ff00 / PhyB-Delta-CreC
                        4: 3446 .. 3502 / #c0c0c0 / P2A
                        5: 3503 .. 4099 / #00ffff / PIF6CreN"
     CDS             596..616
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /note="color: #993366"
                     /translation="PKKKRKV"
     misc_feature    1123
                     /label=silent g>c to abolish SacI site
                     /note="color: #a6acb3"
     misc_feature    1378
                     /label=Silent t>g to abolish HindIII site
                     /note="color: #a6acb3"
     misc_feature    1942
                     /label=silent c>t to abolish MluI Site
                     /note="color: #a6acb3"
     CDS             3446..3502
                     /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."
                     /note="color: #cc99b2
                     Cleavage site after base 3499"
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             3506..3526
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /note="color: #993366"
                     /translation="PKKKRKV"
     polyA_signal    4118..4175
                     /label=Syn PolyA
                     /note="color: #a6acb3"
     polyA_signal    4120..4168
                     /note="synthetic polyadenylation signal"
                     /note="color: #a6acb3"
     primer_bind     complement(4139..4168)
                     /label=SynPA Rev
                     /note="color: black; sequence: 
                     cacacaaaaaaccaacacacagatctaatg; added: 2019-08-14"
     misc_feature    4182..4354
                     /label=ITR
                     /note="color: #c040ff; direction: RIGHT"
     misc_feature    complement(4379..6794)
                     /label=pEMBL8 Plasmid Backbone
                     /note="color: #0000ff; direction: LEFT"
     rep_origin      complement(4588..5176)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(5347..6207)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 5347 .. 6138 / #ccffcc
                        2: 6139 .. 6207 / #ccffcc / signal sequence
                     Cleavage site after base 6138"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVVMVTTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6208..6312)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
     rep_origin      complement(6339..6794)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
ORIGIN
        1 cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc
       61 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc
      121 actaggggtt ccttgtagtt aatgattaac ccgccatgct acttatctac gtagccatgc
      181 tctggaaagc ttcgccctta agctagtatg ccaagtacgc cccctattga cgtcaatgac
      241 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
      301 cagtacatct actcgaggcc acgttctgct tcactctccc catctccccc ccctccccac
      361 ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg gcgaggggcg
      421 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      481 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      541 gggagcggga tcagaatgat ctgatatcat cgatgaattc gagctcgcca ccatgcccaa
      601 gaagaagagg aaggtggttt ccggagtcgg gggtagtggc ggtggccgtg gcggtggccg
      661 tggcggagaa gaagaaccgt cgtcaagtca cactcctaat aaccgaagag gaggagaaca
      721 agctcaatcg tcgggaacga aatctctcag accaagaagc aacactgaat caatgagcaa
      781 agcaattcaa cagtacaccg tcgacgcaag actccacgcc gttttcgaac aatccggcga
      841 atcagggaaa tcattcgact actcacaatc actcaaaacg acgacgtacg gttcctctgt
      901 acctgagcaa cagatcacag cttatctctc tcgaatccag cgaggtggtt acattcagcc
      961 tttcggatgt atgatcgccg tcgatgaatc cagtttccgg atcatcggtt acagtgaaaa
     1021 cgccagagaa atgttaggga ttatgcctca atctgttcct actcttgaga aacctgagat
     1081 tctagctatg ggaactgatg tgagatcttt gttcacttct tccagctcga ttctactcga
     1141 gcgtgctttc gttgctcgag agattacctt gttaaatccg gtttggatcc attccaagaa
     1201 tactggtaaa ccgttttacg ccattcttca taggattgat gttggtgttg ttattgattt
     1261 agagccagct agaactgaag atcctgcgct ttctattgct ggtgctgttc aatcgcagaa
     1321 actcgcggtt cgtgcgattt ctcagttaca ggctcttcct ggtggagata ttaagctgtt
     1381 gtgtgacact gtcgtggaaa gtgtgaggga cttgactggt tatgatcgtg ttatggttta
     1441 taagtttcat gaagatgagc atggagaagt tgtagctgag agtaaacgag atgatttaga
     1501 gccttatatt ggactgcatt atcctgctac tgatattcct caagcgtcaa ggttcttgtt
     1561 taagcagaac cgtgtccgaa tgatagtaga ttgcaatgcc acacctgttc ttgtggtcca
     1621 ggacgatagg ctaactcagt ctatgtgctt ggttggttct actcttaggg ctcctcatgg
     1681 ttgtcactct cagtatatgg ctaacatggg atctattgcg tctttagcaa tggcggttat
     1741 aatcaatgga aatgaagatg atgggagcaa tgtagctagt ggaagaagct cgatgaggct
     1801 ttggggtttg gttgtttgcc atcacacttc ttctcgctgc ataccgtttc cgctaaggta
     1861 tgcttgtgag tttttgatgc aggctttcgg tttacagtta aacatggaat tgcagttagc
     1921 tttgcaaatg tcagagaaac gtgttttgag aacgcagaca ctgttatgtg atatgcttct
     1981 gcgtgactcg cctgctggaa ttgttacaca gagtcccagt atcatggact tagtgaaatg
     2041 tgacggtgca gcatttcttt accacgggaa gtattacccg ttgggtgttg ctcctagtga
     2101 agttcagata aaagatgttg tggagtggtt gcttgcgaat catgcggatt caaccggatt
     2161 aagcactgat agtttaggcg atgcggggta tcccggtgca gctgcgttag gggatgctgt
     2221 gtgcggtatg gcagttgcat atatcacaaa aagagacttt cttttttggt ttcgatctca
     2281 cactgcgaaa gaaatcaaat ggggaggcgc taagcatcat ccggaggata aagatgatgg
     2341 gcaacgaatg catcctcgtt cgtcctttca ggcttttctt gaagttgtta agagccggag
     2401 tcagccatgg gaaactgcgg aaatggatgc gattcactcg ctccagctta ttctgagaga
     2461 ctcttttaaa gaatctggtg gttctgccgg tggctccggt tctggctcca gcggtggcag
     2521 ctctggtgcg tccggcacgg gtactgcggg tggcactggc agcggttccg gtactggctc
     2581 tggcaaccgg aaatggtttc ccgcagaacc tgaagatgtt cgcgattatc ttctatatct
     2641 tcaggcgcgc ggtctggcag taaaaactat ccagcaacat ttgggccagc taaacatgct
     2701 tcatcgtcgg tccgggctgc cacgaccaag tgacagcaat gctgtttcac tggttatgcg
     2761 gcggatccga aaagaaaacg ttgatgccgg tgaacgtgca aaacaggctc tagcgttcga
     2821 acgcactgat ttcgaccagg ttcgttcact catggaaaat agcgatcgct gccaggatat
     2881 acgtaatctg gcatttctgg ggattgctta taacaccctg ttacgtatag ccgaaattgc
     2941 caggatcagg gttaaagata tctcacgtac tgacggtggg agaatgttaa tccatattgg
     3001 cagaacgaaa acgctggtta gcaccgcagg tgtagagaag gcacttagcc tgggggtaac
     3061 taaactggtc gagcgatgga tttccgtctc tggtgtagct gatgatccga ataactacct
     3121 gttttgccgg gtcagaaaaa atggtgttgc cgcgccatct gccaccagcc agctatcaac
     3181 tcgcgccctg gaagggattt ttgaagcaac tcatcgattg atttacggcg ctaaggatga
     3241 ctctggtcag agatacctgg cctggtctgg acacagtgcc cgtgtcggag ccgcgcgaga
     3301 tatggcccgc gctggagttt caataccgga gatcatgcaa gctggtggct ggaccaatgt
     3361 aaatattgtc atgaactata tccgtaacct ggatagtgaa acaggggcaa tggtgcgcct
     3421 gctggaagat ggcgatggaa gcggagctac taacttcagc ctgctgaagc aggctggaga
     3481 cgtggaggag aaccctggac ctatgcccaa gaagaagagg aaggtgatgt tcttaccaac
     3541 cgattattgt tgcaggttaa gcgatcaaga gtatatggag cttgtgtttg agaatggcca
     3601 gattcttgca aagggccaaa gatccaacgt ttctctgcat aatcaacgta ccaaatcgat
     3661 catggatttg tatgaggcag agtataacga ggatttcatg aagagtatca tccatggtgg
     3721 tggtggtgcc atcacaaatc tcggggacac gcaggttgtt ccacaaagtc atgttgctgc
     3781 tgcccatgaa acaaacatgt tggaaagcaa taaacatgtt gacggtggtt ctgccggtgg
     3841 ctccggttct ggctccagcg gtggcagctc tggtgcgtcc ggcacgggta ctgcgggtgg
     3901 cactggcagc ggttccggta ctggctctgg cctgactgtg caccaaaacc tgcctgccct
     3961 ccctgtggat gccacctctg atgaagtcag gaagaacctg atggacatgt tcagggacag
     4021 gcaggccttc tctgaacaca cctggaagat gctcctgtct gtgtgcagat cctgggctgc
     4081 ctggtgcaag ctgaactgag aattctctag actagagcga ataaaagatc tttattttca
     4141 ttagatctgt gtgttggttt tttgtgtgat gcagcacgcg ttacgtagat aagtagcatg
     4201 gcgggttaat cattaactac aaggaacccc tagtgatgga gttggccact ccctctctgc
     4261 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc
     4321 gggcggcctc agtgagcgag cgagcgcgca gctgcattaa tgaatcggcc aacgcgcggg
     4381 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     4441 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     4501 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ccgcagcaaa aggccaggaa
     4561 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     4621 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4681 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     4741 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     4801 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     4861 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4921 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4981 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     5041 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     5101 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     5161 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     5221 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     5281 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     5341 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     5401 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     5461 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     5521 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     5581 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     5641 gcgcaacgtt gttaccatta ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     5701 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     5761 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     5821 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     5881 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     5941 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     6001 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccactatt
     6061 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     6121 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     6181 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     6241 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6301 aggggttccg cgcacatttc cccgaaagat gccacctgaa attataaacg ttaatatttt
     6361 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     6421 cggaaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     6481 ttggaacaag agtccactat tgaggaacgt gaactccagc gtcaaagggc gaaaaaccgt
     6541 ctatcggggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     6601 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     6661 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     6721 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     6781 gctacagggc gcgtcccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     6841 gcgggcctct tcgctattac gc
//
