LOCUS       Exported                8072 bp ds-DNA     circular SYN 23-JUL-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8072)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Jul 24, 2018 from SnapGene Viewer 4.2.1
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8072
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        complement(1..105)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(131..586)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     misc_feature    874..936
                     /label=3' TR
     misc_feature    1113..1344
                     /label=Core Insulator
     misc_feature    1346..1366
                     /label=H1
     misc_feature    1367..1977
                     /label=SH LHA
     misc_feature    1886..1888
                     /label=PAM
     misc_feature    1889..1907
                     /label=SH gRNA 4
     misc_feature    1960..1968
                     /label=SHS231
     misc_feature    1978..2030
                     /label=Bxb1 attP
     promoter        2066..3244
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2297..3235
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     protein_bind    3256..3289
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     CDS             3304..3900
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     misc_feature    3916..3957
                     /label=fC31 attP
     misc_feature    3958..4485
                     /label=SH RHA
     misc_feature    4136..4138
                     /label=PAM
     misc_feature    4139..4157
                     /label=SH gRNA 5
     misc_feature    4486..4506
                     /label=H2
     polyA_signal    4647..4764
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    complement(5021..5251)
                     /label=Core Insulator
     misc_feature    5526..5560
                     /label=5' TR
     primer_bind     complement(6051..6067)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6075..6091
                     /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(6099..6129)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6144..6165
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(6453..7041)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7212..8072)
                     /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 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
       61 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      121 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
      181 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
      241 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
      301 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
      361 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
      421 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
      481 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
      541 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
      601 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
      661 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
      721 cgacgttgta aaacgacggc cagtgagcgc gcctcgttca ttcacgtttt tgaacccgtg
      781 gaggacgggc agactcgcgg tgcaaatgtg ttttacagcg tgatggagca gatgaagatg
      841 ctcgacacgc tgcagaacac gcagctagat taaccctaga aagataatca tattgtgacg
      901 tacgttaaag ataatcatgc gtaaaattga cgcatgtgtt ttatcggtct gtatatcgag
      961 gtttatttat taatttgaat agatattaag ttttattata tttacactta catactaata
     1021 ataaattcaa caaacaattt atttatgttt atttatttat taaaaaaaaa caaaaactca
     1081 aaatttcttc tataaagtaa caaaactttt atgagggaca gccccccccc aaagccccca
     1141 gggatgtaat tacgtccctc ccccgctagg gggcagcagc gagccgcccg gggctccgct
     1201 ccggtccggc gctccccccg catccccgag ccggcagcgt gcggggacag cccgggcacg
     1261 gggaaggtgg cacgggatcg ctttcctctg aacgcttctc gctgctcttt gagcctgcag
     1321 acacctgggg ggatacgggg aaaaggcctc caaggccaga ctagaggtac ccattgagtt
     1381 ctgaggtggt ttgtcataca gcaatggcaa tcagatatat ccacacaaat ataattttag
     1441 tttatatttc tgctactgca gttctcatct tattctgagg atatgtgaca aaataattct
     1501 ttcaaaaata ttgatgctgt gctagattac tattttgaat gaattactag acaaatactt
     1561 catatgtatc tttttttttt tttttttttt tttttttttt tttttttgag acggagtctc
     1621 gctctgtcgc ccaggctgga gtgcagtggc gggatctcgg ctcactgcaa tatgtatctt
     1681 attatgtggg tttacacatt atttatctta ttgatttaac ttcaaaacta aactttagtt
     1741 tagctcttgg gccctatctg gaaagggtca tcttttaatc accattaaat cactgaagtc
     1801 atcagtttat tcaaagtact ctgcacaaat tagcattctt tagtggttgt gaaataaata
     1861 gactttaaac ttatcattaa tattcccaat ggtactatgg gggaggcaaa attttctgtc
     1921 ttcttagtgg tttttttttt ttggctaggg ctaaggatta aactgtcaag cttcgaggtg
     1981 gtttgtctgg tcaaccaccg cggtctcagt ggtgtacggt acaaaccccg agtcgacctt
     2041 ctaggtcttg aaaggagtgg gaattggctc cggtgcccgt cagtgggcag agcgcacatc
     2101 gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaaccggtg cctagagaag
     2161 gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg
     2221 tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt
     2281 tgccgccaga acacaggtaa gtgccgtgtg tggttcccgc gggcctggcc tctttacggg
     2341 ttatggccct tgcgtgcctt gaattacttc cacctggctg cagtacgtga ttcttgatcc
     2401 cgagcttcgg gttggaagtg ggtgggagag ttcgaggcct tgcgcttaag gagccccttc
     2461 gcctcgtgct tgagttgagg cctggcctgg gcgctggggc cgccgcgtgc gaatctggtg
     2521 gcaccttcgc gcctgtctcg ctgctttcga taagtctcta gccatttaaa atttttgatg
     2581 acctgctgcg acgctttttt tctggcaaga tagtcttgta aatgcgggcc aagatctgca
     2641 cactggtatt tcggtttttg gggccgcggg cggcgacggg gcccgtgcgt cccagcgcac
     2701 atgttcggcg aggcggggcc tgcgagcgcg gccaccgaga atcggacggg ggtagtctca
     2761 agctggccgg cctgctctgg tgcctggcct cgcgccgccg tgtatcgccc cgccctgggc
     2821 ggcaaggctg gcccggtcgg caccagttgc gtgagcggaa agatggccgc ttcccggccc
     2881 tgctgcaggg agctcaaaat ggaggacgcg gcgctcggga gagcgggcgg gtgagtcacc
     2941 cacacaaagg aaaagggcct ttccgtcctc agccgtcgct tcatgtgact ccacggagta
     3001 ccgggcgccg tccaggcacc tcgattagtt ctcgagcttt tggagtacgt cgtctttagg
     3061 ttggggggag gggttttatg cgatggagtt tccccacact gagtgggtgg agactgaagt
     3121 taggccagct tggcacttga tgtaattctc cttggaattt gccctttttg agtttggatc
     3181 ttggttcatt ctcaagcctc agacagtggt tcaaagtttt tttcttccat ttcaggtgtc
     3241 gtgaggaatt agcttataac ttcgtatagc atacattata cgaagttatc ctgcagggcc
     3301 accatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc
     3361 gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg
     3421 gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc
     3481 gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg
     3541 gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc
     3601 ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag
     3661 gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg
     3721 ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc
     3781 ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc
     3841 gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc
     3901 tgataatagt ctagagtgcc ccaactgggg taacctttga gttctctcag ttgggggact
     3961 agtgcgtgga caaagagcag taaattgtga aaatggaaca aggcaagggg gcttagacta
     4021 cagtagttaa tcatcaagaa gtgacaaaaa aaataatggt tagttaataa gatttgttta
     4081 agcagatttc tcccagcttt agctctctgt ctctggtgat cagaatgcac tccttccttc
     4141 agactcagtg agcacatctt ccacatggaa gatttcttct ctagctttta ggaaatccag
     4201 agaacccttt ttgtatctgc tgtttttttt tttttaaatg tctttaactc aaaacaattt
     4261 atgtgccagg atgacatatc tttggataat gtgttctgaa ctccttcagt acatacgtat
     4321 ataaattaaa gcaaatattt tttatgataa gctggcataa tagtttcata atttaatcac
     4381 tgatttaaaa atttaattaa aattattttt tgatattttg tgtaataatt tttgaggagt
     4441 atcttttgtg cttaatgagt ggcagatgac acccatgttc ttagctacgc gttaactaag
     4501 taagcggccg ccccggtatc gacaatcaac ctctggatta caaaatttgt gaaagattga
     4561 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     4621 tgtatcatgc gttaactaac taaacttgtt tattgcagct tataatggtt acaaataaag
     4681 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     4741 gtccaaactc atcaatgtat cttatcatgt ctggaattga ctcaaatgat gtcaattagt
     4801 ctatcagaag ctatctggtc tcccttccgg gggacaagac atccctgttt aatatttaaa
     4861 cagcagtgtt cccaaactgg gttcttatat cccttgctct ggtcaaccag gttgcagggt
     4921 ttcctgtcct cacaggaacg aagtccctaa agaaacagtg gcagccaggt ttagccccgg
     4981 aattgactgg attccttttt tagggcccat tggtatggct ttttccccgt atccccccag
     5041 gtgtctgcag gctcaaagag cagcgagaag cgttcagagg aaagcgatcc cgtgccacct
     5101 tccccgtgcc cgggctgtcc ccgcacgctg ccggctcggg gatgcggggg gagcgccgga
     5161 ccggagcgga gccccgggcg gctcgctgct gccccctagc gggggaggga cgtaattaca
     5221 tccctggggg ctttgggggg gggctgtccc tgatatctat aacaagaaaa tatatatata
     5281 ataagttatc acgtaagtag aacatgaaat aacaatataa ttatcgtatg agttaaatct
     5341 taaaagtcac gtaaaagata atcatgcgtc attttgactc acgcggtcgt tatagttcaa
     5401 aatcagtgac acttaccgca ttgacaagca cgcctcacgg gagctccaag cggcgactga
     5461 gatgtcctaa atgcacagcg acggattcgc gctatttaga aagagagagc aatatttcaa
     5521 gaatgcatgc gtcaatttta cgcagactat ctttctaggg ttaatctagc tgcatcagga
     5581 tcatatcgtc gggtcttttt tccggctcag tcatcgccca agctggcgct atctgggcat
     5641 cggggaggaa gaagcccgtg ccttttcccg cgaggttgaa gcggcatgga aagagtttgc
     5701 cgaggatgac tgctgctgca ttgacgttga gcgaaaacgc acgtttacca tgatgattcg
     5761 ggaaggtgtg gccatgcacg cctttaacgg tgaactgttc gttcaggcca cctgggatac
     5821 cagttcgtcg cggcttttcc ggacacagtt ccggatggtc agcccgaagc gcatcagcaa
     5881 cccgaacaat accggcgaca gccggaactg ccgtgccggt gtgcagatta atgacagcgg
     5941 tgcggcgctg ggatattacg tcagcgagga cgggtatcct ggctggatgc cgcagaaatg
     6001 gacatggata ccccgtgagt tacccggcgg gcgcgcttgg cgtaatcatg gtcatagctg
     6061 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     6121 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     6181 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     6241 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     6301 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     6361 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     6421 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     6481 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     6541 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     6601 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     6661 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     6721 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     6781 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     6841 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     6901 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     6961 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     7021 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     7081 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     7141 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     7201 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     7261 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     7321 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     7381 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     7441 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     7501 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     7561 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     7621 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     7681 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     7741 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     7801 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     7861 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     7921 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     7981 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     8041 ataagggcga cacggaaatg ttgaatactc at
//
