LOCUS       sequence-347884-        7380 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR donor plasmid to create GFP fusion proteins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7380)
  TITLE     ENCODE collection: CRISPR constructs
REFERENCE   2  (bases 1 to 7380)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pSTAT5B-donor
FEATURES             Location/Qualifiers
     source          1..7380
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(34..50)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(34..50)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(47..69)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    58..74
                     /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(join(82..88,89..106,107..112))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    127..148
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(265..282)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(436..1024)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(516..535)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1195..1986,1987..2055))
                     /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     1818..1837
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2056..2160)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2228..2246
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2284..2306)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2406..2425
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2619..2641
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2633..2650
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2634..2650
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3873..3893
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3906..3950
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             3990..4013
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(4041..4043,4044..4046,4047..4760)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVRLADHYQQDTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLK
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4086..4107)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4347..4366)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    join(4798..5371,5372..5374,5375..5383)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4964..4981)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5191..5210
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5465..6268
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5528..5547)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6138..6157
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 gacctgcagg catgcaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
       61 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
      121 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
      181 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
      241 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
      301 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
      361 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
      421 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
      481 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
      541 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
      601 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
      661 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
      721 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
      781 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
      841 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
      901 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
      961 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     1021 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     1081 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     1141 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     1201 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     1261 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     1321 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     1381 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     1441 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     1501 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     1561 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     1621 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     1681 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     1741 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     1801 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     1861 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     1921 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     1981 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     2041 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     2101 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     2161 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     2221 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     2281 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     2341 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     2401 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
     2461 cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg
     2521 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     2581 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
     2641 gacggccagt gaattcgagc tcggtacccc cttagataag ttaggtggtt gtgttctgtc
     2701 ctaagaaaag atgtgattca taaggaagat cacatttttg tttcacttga gaactttgat
     2761 ttactacatt cttcccttgg cagctaaagc tgttgatgga tacgtgaagc cacagatcaa
     2821 gcaagtggtc cctgagtaag tgtccaggtg gctgtggctc tccttctgcc tctttcctcc
     2881 tcccccagac cctgcctccc atccttatcc tggcccagct ttccgctccc ccggggaaac
     2941 tgtctcagcc ctggggaagg acctgggctt ggtgatgcta tgccagtacc tgaggcccca
     3001 ctcctgtagc tggagcccca gggccgagtg gggcactttg tccttctaca ccctgagaaa
     3061 gacaaagata ccccttggtc ccctccccaa gtcagctgcc ctggcctccc tggcagctga
     3121 agctctgttc tcttccttct gcaggtttgt gaacgcatct gcagatgccg ggggcggcag
     3181 cgccacgtac atggaccagg ccccctcccc agctgtgtgt ccccaggctc actataacat
     3241 gtacccacag aagtaggtgg cattcttgtg ggggtccaca ggggaggaac tgggggcttg
     3301 gccccaggct ggacccctgg agggctgggg tcaaaggaat ccagagctcc tctaggtctg
     3361 agaccacaca gccctggccc gggctctggc ctttgggagg gagagatttg accagaagtg
     3421 attggtggtt tagagtcggg ttgggcaaat gtcatctgtt cccctctctc aatatggaga
     3481 tttttcttgc tctaaggact gagtggagtt gtgatgtgtc tggacagccg gtgtgagagg
     3541 taacagggga gggtatttca gtctgagttg ctgtacagcc agctgtttga atggttgttc
     3601 ttttcccatg aagccccagg ctgagggcat aggctggctc agactgctcc cctggtggcc
     3661 tgtggggctt ggggtggtct ctggcaggct ttccccttcc cttcatccct cttctttccc
     3721 tcccaagccc tgactcagtc cttgacaccg atggggactt cgatctggag gacacaatgg
     3781 acgtagcgcg gcgtgtggag gagctcctgg gccggccaat ggacagtcag tggatcccgc
     3841 acgcacaatc ggggattcca actactgcaa gcgagaatct ttattttcag ggcgccgcca
     3901 aattcaaaga aaccgctgct gctaaattcg aacgccagca catggacagc ggaggtggag
     3961 gttcgagcgg tccctcgggt tcgtcgagcc tggaagttct gttccagggg cccctgtcgt
     4021 cgagcggtcc ctcgggttcg atggtgagca aaggcgagga gctgttcacc ggggtggtgc
     4081 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     4141 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     4201 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
     4261 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     4321 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     4381 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     4441 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     4501 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     4561 acggcagcgt gcggctcgcc gaccactacc agcaggacac ccccatcggc gacggccccg
     4621 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
     4681 agaagcgcga tcacatggtc cttaaggagt tcgtgaccgc cgccgggatc actctcggca
     4741 tggacgagct gtacaagtaa tgatctagag tcgagttaat taagaattcc gccccccccc
     4801 ctctccctcc ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg
     4861 tttgtctata tgttattttc caccatattg ccgtcttttg gcaatgtggg ggcccggaaa
     4921 cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg
     4981 cagggtctgt tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca
     5041 acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc
     5101 ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt
     5161 gtgagttgga tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg
     5221 ctgaaggatg cccagaaggt accccattgt atgggatctg atctggggcc tcggtgcaca
     5281 tgctttacat gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa ccacggggac
     5341 gtggttttcc tttgaaaaac acgatgataa tatggccaca acctgttaca ttgcacaaga
     5401 taaaaatata tcatcacgaa cagtaaaact gtctgcttac ataaacagta atacaagggg
     5461 tgttatggga tcggccattg aacaggatgg attgcacgca ggttctccgg ccgcttgggt
     5521 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     5581 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     5641 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     5701 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     5761 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     5821 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     5881 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     5941 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     6001 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     6061 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     6121 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     6181 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     6241 ctatcgcctt cttgacgagt tcttctgagt cccccgcgac ctctccatct tcagcttctt
     6301 catcttcacc agaggaatca ctcttgtgga tgttttaatt ccatgaatcg cttctctttt
     6361 gaaacaatac tcataatgtg aagtgttaat actagttgtg accttagtgt ttctgtgcat
     6421 ggtggcacca gcgaagggag tgcgagtatg tgtttgtgtg tgtgtgtgtg tgtgtgtgtg
     6481 cgtgtttgca cgttatggtg tttctccctc tcactgtctg agagtttagt tgtagcagag
     6541 gggccacaga cagaagctgt ggtggttttt actttgtgca aaaaggcagt gagtttcgtg
     6601 aagcctggaa gttggtcatg tgtcttaaga gtggctggac tttgacatgt ggctgtttga
     6661 ataagagaag gacaaaggga ggagaaagca catgtgctcc agtgagtctt cgtcactctg
     6721 tctgccaagc aattgatata taaccgtgat tgtctctgct tttcttctga aatgtagata
     6781 actgcttttt gacaaagaga gccttccctc tcccccaccc ctgtgttctt gggtaggaat
     6841 gggaaaaggg gcaacctaca aagattgttg gggcaaggga agtcacaagc tttcggatgg
     6901 gcggtggctt ttcacaaaac atttagctca tcttattctc tctttgtcct ctctcccctc
     6961 ctgcccgccc gcaccctgga attgccactc agttcctctg ggtgtgcaca tatgtttgga
     7021 gaaatagagg agagaaaaga gggccacgta actgagagct tacagtgcca atgccgtttg
     7081 tgttctggcc agagtggagt gcgcagccct gactcccagg cgctgagatc gttgcctggt
     7141 tacccaggaa gctgctgttc cggctgccca gcccttctct gagccagcgg atgcacagtc
     7201 cgtggccttc ttcaggctta ttgatgatgc tttttgcaaa tgttgaatca tggttctgtt
     7261 tctaagttgg atcttttttg ttttctcctt gccaccctaa tttgacatca aaattctctc
     7321 ttgtgcattg ggccctgggt cattcaaacc caggtcacct cattcccctt ctctgttcac
//
