LOCUS       sequence-353735-        5897 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SyntheticTranscriptionFactor_loxFAS_PGK_MCS_scarlett_ERD_ZIFDBD_P2A_
            Neo_loxP_AMP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5897)
  AUTHORS   Staller MV, Ramirez E, Kotha SR, Holehouse AS, Pappu RV, Cohen BA
  TITLE     Directed mutational scanning reveals a balance between acidic and 
            hydrophobic residues in strong human activation domains.
  JOURNAL   Cell Syst. 2022 Apr 20;13(4):334-345.e5. doi: 
            10.1016/j.cels.2022.01.002. Epub 2022 Feb 3.
  PUBMED    35120642
REFERENCE   2  (bases 1 to 5897)
  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"; doi: 
            "10.1016/j.cels.2022.01.002"; journalName: "Cell Syst"; date: 
            "2022-04-20- 20"; volume: "13"; issue: "4"; pages: "334-345.e5"
COMMENT     Sequence Label: pMVS223
FEATURES             Location/Qualifiers
     source          1..5897
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        74..584
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     503..521
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     regulatory      590..599
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             611..1306
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(744..762)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1012..1031
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             2741..2797
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             2801..3601
                     /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="GSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQG
                     RPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                     LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEE
                     HQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQD
                     IALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2861..2880)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3471..3490
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     protein_bind    3623..3656
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(3754..3771)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3925..4513)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4005..4024)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4684..5475,5476..5544))
                     /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     5307..5326
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5545..5649)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5717..5735
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5773..5795)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     join(5895..5897,1..17)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 catcagagca gattgtactg agagtgcaca caacttcgta tatacctttc tatacgaagt
       61 tgtgaattcc cacggggttg gggttgcgcc ttttccaagg cagccctggg tttgcgcagg
      121 gacgcggctg ctctgggcgt ggttccggga aacgcagcgg cgccgaccct gggtctcgca
      181 cattcttcac gtccgttcgc agcgtcaccc ggatcttcgc cgctaccctt gtgggccccc
      241 cggcgacgct tcctgctccg cccctaagtc gggaaggttc cttgcggttc gcggcgtgcc
      301 ggacgtgaca aacggaagcc gcacgtctca ctagtaccct cgcagacgga cagcgccagg
      361 gagcaatggc agcgcgccga ccgcgatggg ctgtggccaa tagcggctgc tcagcggggc
      421 gcgccgagag cagcggccgg gaaggggcgg tgcgggaggc ggggtgtggg gcggtagtgt
      481 gggccctgtt cctgcccgcg cggtgttccg cattctgcaa gcctccggag cgcacgtcgg
      541 cagtcggctc cctcgttgac cgaatcaccg acctctctcc ccagcttaag ccaccatggg
      601 atccggcagc atggtgagca agggcgaggc agtgatcaag gagttcatgc ggttcaaggt
      661 gcacatggag ggctccatga acggccacga gttcgagatc gagggcgagg gcgagggccg
      721 cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt
      781 ctcctgggac atcctgtccc ctcagttcat gtacggctcc agggccttca ccaagcaccc
      841 cgccgacatc cccgactact ataagcagtc cttccccgag ggcttcaagt gggagcgcgt
      901 gatgaacttc gaggacggcg gcgccgtgac cgtgacccag gacacctccc tggaggacgg
      961 caccctgatc tacaaggtga agctccgcgg caccaacttc cctcctgacg gccccgtaat
     1021 gcagaagaag acaatgggct gggaagcgtc caccgagcgg ttgtaccccg aggacggcgt
     1081 gctgaagggc gacattaaga tggccctgcg cctgaaggac ggcggccgct acctggcgga
     1141 cttcaagacc acctacaagg ccaagaagcc cgtgcagatg cccggcgcct acaacgtcga
     1201 ccgcaagttg gacatcacct cccacaacga ggactacacc gtggtggaac agtacgaacg
     1261 ctccgagggc cgccactcca ccggcggcat ggacgagctg tacaagggca gcgctggctc
     1321 tgctggagac atgagagctg ccaacctttg gccaagcccg ctcatgatca aacgctctaa
     1381 gaagaacagc ctggccttgt ccctgacggc cgaccagatg gtcagtgcct tgttggatgc
     1441 tgagcccccc atactctatt ccgagtatga tcctaccaga cccttcagtg aagcttcgat
     1501 gatgggctta ctgaccaacc tggcagacag ggagctggtt cacatgatca actgggcgaa
     1561 gagggtgcca ggctttgtgg atttgaccct ccatgatcag gtccaccttc tagaatgtgc
     1621 ctggctagag atcctgatga ttggtctcgt ctggcgctcc atggagcacc cagtgaagct
     1681 actgtttgct cctaacttgc tcttggacag gaaccaggga aaatgtgtag agggcatggt
     1741 ggagatcttc gacatgctgc tggctacatc atctcggttc cgcatgatga atctgcaggg
     1801 agaggagttt gtgtgcctca aatctattat tttgcttaat tctggagtgt acacatttct
     1861 gtccagcacc ctgaagtctc tggaagagaa ggaccatatc caccgagtcc tggacaagat
     1921 cacagacact ttgatccacc tgatggccaa ggcaggcctg accctgcagc agcagcacca
     1981 gcggctggcc cagctcctcc tcatcctctc ccacatcagg cacatgagta acaaaggcat
     2041 ggagcatctg tacagcatga agtgcaagaa cgtggtgccc ctctatgacc tgctgctgga
     2101 gatgctggac gcccaccgcc tacatgcgcc cactagccgt ggaggggcat ccgtggagga
     2161 gacggaccaa agccacttgg ccactgcggg ctctacttca tcgggtacct ctagaccagg
     2221 cgaacgaccg tttcaatgcc ggatatgtat gaggaacttc tccgaggagg caaacttgag
     2281 gcgccacacc cgaacacata caggagaaaa gccattccaa tgtcgaattt gtatgcgcaa
     2341 tttttcagat cactcaagcc tcaagcgaca cctccgcaca catactggtt cacagaagcc
     2401 ctttcagtgc aggatttgta tgcgcaactt tagccaatca gcgaaccttt tgcggcacac
     2461 tagaacgcat acaggtgaga agcctttcca gtgtcgcatc tgtatgcgga acttcagcga
     2521 ccccagttca ttgaagaggc atttgcgaac tcacaccggt tctcaaaaac cttttcagtg
     2581 ccgaatttgt atgcgcaact tcagccaaca aacaaatttg acgagacaca cgcgcacgca
     2641 caccggggaa aaaccgtttc aatgtcgaat ctgcatgcgc aattttagcg atgcgacaca
     2701 acttgttagg catctgcgca cacacttgcg gggaagcgga gctactaact tcagcctgct
     2761 gaagcaggct ggagacgtgg aggagaaccc tggacctgca ggatcggcca ttgaacaaga
     2821 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     2881 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     2941 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     3001 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     3061 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     3121 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     3181 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     3241 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     3301 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg atgatctcgt
     3361 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     3421 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     3481 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     3541 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     3601 agctagcatc gatcgactcg agataacttc gtataatgta tgctatacga agttataatt
     3661 gaatctgatt tcgataagcc aggttaacct gcattaatga atcggccaac gcgcggggag
     3721 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     3781 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     3841 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     3901 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     3961 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     4021 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4081 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4141 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     4201 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     4261 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     4321 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     4381 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     4441 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     4501 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     4561 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     4621 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     4681 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     4741 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     4801 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     4861 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     4921 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     4981 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     5041 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     5101 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     5161 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     5221 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     5281 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     5341 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     5401 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     5461 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     5521 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     5581 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     5641 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     5701 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
     5761 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     5821 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     5881 ctggcttaac tatgcgg
//
