LOCUS       Exported                3488 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  human GFP11 with tagRFP 33bp downstream in Lamin A/C (recoded).
ACCESSION   .
VERSION     .
KEYWORDS    pAP1893-1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3488)
  AUTHORS   Paix A, Folkmann A, Goldman DH, Kulaga H, Grzelak MJ, Rasoloson D, 
            Paidemarry S, Green R, Reed RR, Seydoux G
  TITLE     Precision genome editing using synthesis-dependent repair of 
            Cas9-induced DNA breaks.
  JOURNAL   Proc Natl Acad Sci U S A. 2017 Nov 28. pii: 1711979114. doi: 
            10.1073/pnas.1711979114.
  PUBMED    29183983
REFERENCE   2  (bases 1 to 3488)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3488
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     105..124
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     318..340
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     332..349
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     333..349
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             376..423
                     /codon_start=1
                     /product="11th beta-strand of superfolder GFP"
                     /label=GFP11
                     /note="associates with GFP(1-10), consisting of the first 
                     10 beta-strands, to reconstitute fluorescent GFP (Cabantous
                     et al., 2005; Kamiyama et al., 2016)"
                     /translation="RDHMVLHEYVNAAGIT"
     primer_bind     complement(1221..1237)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1221..1237)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1234..1256)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1245..1261
                     /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(1269..1299)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1314..1335
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1452..1469)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1623..2211)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1703..1722)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2382..3242)
                     /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     3005..3024
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3243..3347)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3415..3433
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3471..5)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
       61 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
      121 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
      181 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
      241 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
      301 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattcgagct
      361 cggtacccgg ggatccgtga ccacatggtc cttcatgagt atgtaaatgc tgctgggatt
      421 acaggtggcg gccagcgccg tgctacgcga agtggagctc aagcggtctc taaaggggaa
      481 gaactcatca aggagaatat gcacatgaag ctctatatgg aaggtacagt aaacaatcac
      541 cacttcaaat gcacttctga aggtgaagga aagccctacg agggtacaca gacgatgaga
      601 attaaagtcg ttgagggagg cccgctgccc tttgcttttg acatccttgc aacctcattt
      661 atgtatggca gccgaacatt catcaatcat acacagggga ttccagattt tttcaaacag
      721 tcctttccgg agggctttac atgggagcga gtaacgacct atgaagatgg gggagttttg
      781 accgccacgc aagatacgtc actccaagac ggctgcctga tttacaatgt aaaaataaga
      841 ggcgttaatt ttccgtctaa tggcccggta atgcagaaaa agaccctggg ttgggaggcc
      901 aacacggaaa tgctttatcc ggcagacggt ggcttggagg ggcgctctga catggcactg
      961 aagttggtcg gcggaggtca ccttatttgc aacttcaaaa ctacgtaccg ctcaaagaag
     1021 ccagccaaaa accttaaaat gcccggggtt tattatgtag accaccgcct cgaacggatc
     1081 aaggaagctg ataaagaaac gtatgtagag caacatgaag tcgcagtggc aaggtactgt
     1141 gatctcccca gtaagctggg gcacaaactg aatgatcctc tagagtcgac ctgcaggcat
     1201 gcaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     1261 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     1321 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     1381 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     1441 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     1501 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     1561 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     1621 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     1681 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     1741 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     1801 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     1861 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     1921 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     1981 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     2041 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     2101 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     2161 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     2221 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     2281 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     2341 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     2401 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     2461 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     2521 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     2581 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     2641 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     2701 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     2761 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     2821 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     2881 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     2941 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     3001 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     3061 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     3121 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     3181 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     3241 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     3301 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     3361 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     3421 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     3481 atgcagct
//
