LOCUS       Exported                4176 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Synthetic promoter responsive to gRNA2.
ACCESSION   .
VERSION     .
KEYWORDS    P2-ECFP-pA (Construct 6)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4176)
  AUTHORS   Nissim L, Perli SD, Fridkin A, Perez-Pinera P, Lu TK
  TITLE     Multiplexed and Programmable Regulation of Gene Networks with an 
            Integrated RNA and CRISPR/Cas Toolkit in Human Cells.
  JOURNAL   Mol Cell. 2014 May 14. pii: S1097-2765(14)00355-4. doi: 
            10.1016/j.molcel.2014.04.022.
  PUBMED    24837679
REFERENCE   2  (bases 1 to 4176)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4176
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             complement(448..1308)
                     /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     1071..1090
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1309..1413)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1440..1895
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1527..1546)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1737..1758
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    2087..2178
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     2127..2146
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     CDS             2575..3294
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2620..2641)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2881..2900)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3228..3249
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3320..3441
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3331..3350)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3385..3404
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3694..3711)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3865..277)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3945..3964)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
       61 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
      121 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
      181 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
      241 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
      301 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
      361 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
      421 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
      481 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
      541 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
      601 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
      661 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
      721 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
      781 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
      841 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
      901 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
      961 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     1021 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     1081 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     1141 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     1201 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     1261 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     1321 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     1381 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     1441 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     1501 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     1561 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     1621 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     1681 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     1741 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     1801 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     1861 cgcgaatttt aacaaaatat taacgcttac aatttgccat tcgccattca ggctgcgcaa
     1921 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagccca agctaccatg
     1981 ataagtaagt aatattaagg tacgggaggt acttggagcg gccgcaataa atatctttat
     2041 tttcattaca tctgtgtgtt ggttttttgt gtgaatcgat agtactaaca tacgctctcc
     2101 atcaaaacaa aacgaaacaa aacaaactag caaaataggc tgtccccagt gcaagtgcag
     2161 gtgccagaac atttctctat cgatagctag cccaggacag tactccgact tacttaatat
     2221 tttcaggcta gcccaggaca gtactccgac ttacttaata ttttcaggct agcccaggac
     2281 agtactccga cttacttaat attttcaggc tagcccagga cagtactccg acttacttaa
     2341 tattttcagg ctagcccagg acagtactcc gacttactta atattttcag gctagcccag
     2401 gacagtactc cgacttactt aatattttca ggctagccca ggacagtact ccgactaact
     2461 taatattttc aggctagccc aggacagtac tccgacttac taatattttc agctagcggg
     2521 gggctataaa agggggtggg ggcgttcgtc ctgctatcta gcgtcgcgtt gaccatggtg
     2581 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     2641 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
     2701 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     2761 accaccctga cctggggcgt gcagtgcttc gcccgctacc ccgaccacat gaagcagcac
     2821 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     2881 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     2941 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     3001 gagtacaacg ccatcagcga caacgtctat atcaccgccg acaagcagaa gaacggcatc
     3061 aaggccaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     3121 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     3181 agcacccagt ccaagctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     3241 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaaggccgg
     3301 ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     3361 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     3421 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     3481 ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg taaaatcgat
     3541 aaggatcgag tatgtcgacc gatgcccttg agagccttca acccagtcag ctccttccgg
     3601 tgggcgcggg gcatgactat cgtcgccgca cttatgactg tcttctttat catgcaactc
     3661 gtaggacagg tgccggcagc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     3721 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     3781 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     3841 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     3901 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     3961 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4021 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4081 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     4141 cgaccgctgc gccttatccg gtaactatcg tcttga
//
