LOCUS       Exported                5341 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  synthetic circuit: AND gate.
ACCESSION   .
VERSION     .
KEYWORDS    pAN-AND
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5341)
  AUTHORS   Nielsen AA, Voigt CA
  TITLE     Multi-input CRISPR/Cas genetic circuits that interface host 
            regulatory networks.
  JOURNAL   Mol Syst Biol. 2014 Nov 24;10:763. doi: 10.15252/msb.20145735.
  PUBMED    25422271
REFERENCE   2  (bases 1 to 5341)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5341
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      complement(169..757)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(249..268)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(928..1788)
                     /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"
     primer_bind     1551..1570
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1789..1893)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      2971..3042
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3058..3085
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_RNA        3182..3257
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    3412..3433
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             complement(3434..4312)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        4339..4623
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     4546..4565
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     misc_RNA        4644..4719
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             4740..4769
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(4855..4872)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      5025..5071
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
       61 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
      121 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
      181 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
      241 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
      301 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
      361 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
      421 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
      481 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
      541 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
      601 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
      661 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
      721 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
      781 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
      841 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
      901 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
      961 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     1021 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     1081 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     1141 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     1201 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     1261 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     1321 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     1381 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     1441 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     1501 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     1561 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     1621 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     1681 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     1741 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     1801 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     1861 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     1921 cgtctaagaa accattatta tcatgacatt aaccagttgc tttccgaatg acatgcgtct
     1981 cctttacacc tccacaacta gctggtatta tgctagcata atacctagga ctgagctgtt
     2041 ttagagctag aaatagcaag ttaaaataag gctagtccga ttgcaacttg aaaaagtggc
     2101 accgagtcgg tgcttttttt ctcggtacca aattccagaa aagagacgct ttcgagcgtc
     2161 ttttttcgtt ttggtccgat aagtccctaa cttttacagc tagctcagtc ctaggtatta
     2221 tgctagcctg aagtacgtct gagcgtgata cccgctcact gaagatgtcc cggtagggcc
     2281 gaaacgtacc tctacaaata attttgttta actatggact atgtttgaaa gggagaaata
     2341 ctagatggca cgtaccccga gccgtagcag cattggtagc ctgcgtagtc cgcataccca
     2401 taaagcaatt ctgaccagca ccattgaaat cctgaaagaa tgtggttata gcggtctgag
     2461 cattgaaagc gttgcacgtc gtgccggtgc aagcaaaccg accatttatc gttggtggac
     2521 caataaagca gcactgattg ccgaagtgta tgaaaatgaa agcgaacagg tgcgtaaatt
     2581 tccggatctg ggtagcttta aagccgatct ggattttctg ctgcgtaatc tgtggaaagt
     2641 ttggcgtgaa accatttgtg gtgaagcatt tcgttgtgtt attgcagaag cacagctgga
     2701 ccctgcaacc ctgacccagc tgaaagatca gtttatggaa cgtcgtcgtg agatgccgaa
     2761 aaaactggtt gaaaatgcca ttagcaatgg tgaactgccg aaagatacca atcgtgaact
     2821 gctgctggat atgatttttg gtttttgttg gtatcgcctg ctgaccgaac agctgaccgt
     2881 tgaacaggat attgaagaat ttaccttcct gctgattaat ggtgtttgtc cgggtacaca
     2941 gcgttaacta gggcccatac ccccaggcat caaataaaac gaaaggctca gtcgaaagac
     3001 tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tctactagag tcacactggc
     3061 tcaccttcgg gtgggccttt ctgcgtttat atacgcgacg tacggtggaa tctgattcgt
     3121 taccaattga catgatacga aacgtaccgt atcgttaagg tataatacca gctagttgtg
     3181 ggttttagag ctagaaatag caagttaaaa taaggctagt ccgttatcaa cttgaaaaag
     3241 tggcaccgag tcggtgcttt ttttctcggt accaaagacg aacaataaga cgctgaaaag
     3301 cgtctttttt cgttttggtc cttcagccaa aaaacttaag accgccggtc ttgtccacta
     3361 ccttgcagta atgcggtgga caggatcggc ggttttcttt tctcttctca agaattcgcg
     3421 gccgcttcta gagttatgac aacttgacgg ctacatcatt cactttttct tcacaaccgg
     3481 cacggaactc gctcgggctg gccccggtgc attttttaaa tacccgcgag aaatagagtt
     3541 gatcgtcaaa accaacattg cgaccgacgg tggcgatagg catccgggtg gtgctcaaaa
     3601 gcagcttcgc ctggctgata cgttggtcct cgcgccagct taagacgcta atccctaact
     3661 gctggcggaa aagatgtgac agacgcgacg gcgacaagca aacatgctgt gcgacgctgg
     3721 cgatatcaaa attgctgtct gccaggtgat cgctgatgta ctgacaagcc tcgcgtaccc
     3781 gattatccat cggtggatgg agcgactcgt taatcgcttc catgcgccgc agtaacaatt
     3841 gctcaagcag atttatcgcc agcagctccg aatagcgccc ttccccttgc ccggcgttaa
     3901 tgatttgccc aaacaggtcg ctgaaatgcg gctggtgcgc ttcatccggg cgaaagaacc
     3961 ccgtattggc aaatattgac ggccagttaa gccattcatg ccagtaggcg cgcggacgaa
     4021 agtaaaccca ctggtgatac cattcgcgag cctccggatg acgaccgtag tgatgaatct
     4081 ctcctggcgg gaacagcaaa atatcacccg gtcggcaaac aaattctcgt ccctgatttt
     4141 tcaccacccc ctgaccgcga atggtgagat tgagaatata acctttcatt cccagcggtc
     4201 ggtcgataaa aaaatcgaga taaccgttgg cctcaatcgg cgttaaaccc gccaccagat
     4261 gggcattaaa cgagtatccc ggcagcaggg gatcattttg cgcttcagcc atacttttca
     4321 tactcccgcc attcagagaa gaaaccaatt gtccatattg catcagacat tgccgtcact
     4381 gcgtctttta ctggctcttc tcgctaacca aaccggtaac cccgcttatt aaaagcattc
     4441 tgtaacaaag cgggaccaaa gccatgacaa aaacgcgtaa caaaagtgtc tataatcacg
     4501 gcagaaaagt ccacattgat tatttgcacg gcgtcacact ttgctatgcc atagcatttt
     4561 tatccataag attagcggat cctacctgac gctttttatc gcaactctct actgtttctc
     4621 catataatac ccgtgtgacc cgtgttttag agctagaaat agcaagttaa aataaggcta
     4681 gtccgttatc aacttgaaaa agtggcaccg agtcggtgct ttttttgaag cttgggcccg
     4741 aacaaaaact catctcagaa gaggatctga atagcgccgt cgaccatcat catcatcatt
     4801 gagtttaaac ggtctccagc ttggctgttt tggcggatga gagaagattt tcagcctgat
     4861 acagattaaa tcagaacgca gaagcggtct gataaaacag aatttgcctg gcggcagtag
     4921 cgcggtggtc ccacctgacc ccatgccgaa ctcagaagtg aaacgccgta gcgccgatgg
     4981 tagtgtgggg tctccccatg cgagagtagg gaactgccag gcatcaaata aaacgaaagg
     5041 ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac tggatcctta
     5101 ctcgagtcta gactgcaggc ttcctcgata ggaagcagga atagttttta caccaacggg
     5161 tcacacgggt attatgctag cataatacct aggactgagc tgttttagag ctagaaatag
     5221 caagttaaaa taaggctagt ccggcttcaa cttgaaaaag tggcaccgag tcggtgcttt
     5281 ttttggaaac acagaaaaaa gcccgcacct gacagtgcgg gctttttttt tcgaccaaag
     5341 g
//
