LOCUS       Exported                3876 bp ds-DNA     circular SYN 16-DEC-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3876)
  AUTHORS   Rob
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Feb 25, 2021 from SnapGene Viewer 5.2.4
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3876
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          714..1679
                     /organism="pGEM-T Easy Vector"
                     /mol_type="other DNA"
     source          3454..3829
                     /organism="pYPQ132A"
                     /mol_type="other DNA"
     promoter        17
                     /label=T7_promoter
                     /note="/translation=MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLG
                     ARVGY 
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE 
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL 
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL 
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA 
                     EIGASLIKHW*"
     primer_bind     complement(24..40)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      181..636
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        749..777
                     /label=AmpR_promoter
                     /note="/translation=MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLG
                     ARVGY 
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE 
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL 
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL 
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA 
                     EIGASLIKHW*"
     gene            819..1679
                     /gene="Ampicillin"
                     /label=Ampicillin
     CDS             819..1679
                     /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"
     misc_feature    1535
                     /label=mutation
     rep_origin      1850..2438
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2824..2840
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2858..2876
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     misc_feature    2950..3352
                     /label=pU3 promoter
     misc_signal     complement(3355..3360)
                     /label=BbsI
     misc_signal     3363..3368
                     /label=BbsI
                     /label=BbsI(1)
     misc_RNA        3371..3446
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    3447..3453
                     /label=Termination signal
     regulatory      3835..3844
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     promoter        complement(join(3875..3876,1..17))
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 tatagtgagt cgtattacaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac
       61 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
      121 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
      181 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
      241 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
      301 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
      361 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
      421 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
      481 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
      541 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
      601 acgcgaattt taacaaaata ttaacgctta caatttcctg atgcggtatt ttctccttac
      661 gcatctgtgc ggtatttcac accgcatcag gtggcacttt tcggggaaat gtgcgcggaa
      721 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
      781 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
      841 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
      901 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
      961 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     1021 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     1081 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     1141 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     1201 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     1261 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     1321 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     1381 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     1441 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     1501 ttattgctga taaatctgga gccggtgagc gtggatctcg cggtatcatt gcagcactgg
     1561 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     1621 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     1681 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     1741 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     1801 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     1861 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     1921 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     1981 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     2041 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     2101 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     2161 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     2221 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     2281 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     2341 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     2401 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     2461 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     2521 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     2581 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
     2641 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     2701 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg
     2761 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
     2821 acacaggaaa cagctatgac catgattacg ccaagctatt taggtgacac tatagaatac
     2881 tcaagctatg catccaacgc gttgggagct ctcccatatg gtcgacctgc aggcggccgg
     2941 tctcccatgc gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc
     3001 atgcctcctc tagaggtgtc gctattttac tttaaatttt tcttatggct cagcctgtga
     3061 tggataactg aatcaaacaa atggcgtctg ggtttaagaa catctgtttt ggctatgttg
     3121 gacgaaacaa gtgaactttt aggatcaact tccgtttata tacggagctt atatcgagca
     3181 ataagataag tgggcttttt atgtaattta atgggctatc gtccatatat tcactaatac
     3241 ccatgcccag tacccatgta tgcgtttcat ataagctcct aatttctccc acatcgctca
     3301 aatctaaaca aatcttgttg tatatataac actgagggag caccattggt cagggtcttc
     3361 gagaagacct gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac
     3421 ttgaaaaagt ggcaccgagt cggtgctttt tttgcaaaaa ttttcagatt ttttcttcat
     3481 ctgtagattt ctgggttttt ttttccgttt cgtgaatcat aagtgaagtt ttggatgcaa
     3541 atctgcgcga aaaaagttgg acctgcaatg agcttattta gatagctaag acaaagtgat
     3601 tggtccgttg tttcagttct gattgtcaga gagtttgttt cgagtcggcg acaccaatgc
     3661 gttttgttaa ccagatttcg ggtaagaaat gtatcgagag tttgtttcca gacggctaca
     3721 tcattttctt atgaagggtg aaattagata gaccaaagat tgaaacacaa catttctttc
     3781 acaaaaatat aataaacttg atagcattta ggatcagcgg accgagaccg gccgccatgg
     3841 cggccgggag catgcgacgt cgggcccaat tcgccc
//
