LOCUS       sequence-323916-        4718 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EYFP reporter vector for the ZF43x12-Spaced-Far promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4718)
  AUTHORS   Donahue PS, Draut JW, Muldoon JJ, Edelstein HI, Bagheri N, Leonard 
            JN
  TITLE     The COMET toolkit for composing customizable genetic programs in 
            mammalian cells.
  JOURNAL   Nat Commun. 2020 Feb 7;11(1):779. doi: 10.1038/s41467-019-14147-5.
  PUBMED    32034124
REFERENCE   2  (bases 1 to 4718)
  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"; journalName: "Nat 
            Commun."; date: "2020-02-7"; volume: "11(1):779. doi"; pages: " 
            10.1038/s41467-019-14147-5"
COMMENT     Sequence Label: pPD150 ZF43x12-Spaced-Far_EYFP
FEATURES             Location/Qualifiers
     source          1..4718
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      578..587
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(584..586,587..589,590..1303)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(629..650)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(890..909)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1237..1258
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1348..1365)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1354..1578
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1624..2052
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1711..1730)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1921..1942
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2075..2271
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2122..2257
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2184..2203
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    2391..2512
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2428..2447)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2482..2501
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2561..2577)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2561..2577)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2574..2596)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2585..2601
                     /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(2609..2615,2616..2633,2634..2639))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2654..2675
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2792..2809)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2963..3551)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3043..3062)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3722..4513,4514..4582))
                     /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     4345..4364
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4583..4687)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctgaa ttcggatcca gagtgaggac tacctataga agtaagagtg
       61 aggacaatcg tatacaagta gatccagagt gaggactacc tatagaagta agagtgagga
      121 caatcgtata caagtagatc cagagtgagg actacctata gaagtaagag tgaggacaat
      181 cgtatacaag tagatccctc tagagggaga tccagagtga ggactaccta tagaagtaag
      241 agtgaggaca atcgtataca agtagatcca gagtgaggac tacctataga agtaagagtg
      301 aggacaatcg tatacaagta gatccagagt gaggactacc tatagaagta agagtgagga
      361 caatcgtata caagtagatc cctctagagg gagatcctga gtagtgcgcg agcaaaattt
      421 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc
      481 gttttgcgct gcttcgcgat gtacgggcca gatatacgat ccctctagag ggtatataat
      541 gggggccact agtctactac cagagctcat cgctagcgcc accatggtga gcaagggcga
      601 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
      661 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
      721 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccttcgg
      781 ctacggcctg cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa
      841 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
      901 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
      961 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     1021 caacagccac aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt
     1081 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
     1141 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gctaccagtc
     1201 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     1261 cgccgccggg atcactctcg gcatggacga gctgtacaag taggcggccg ctcgagtcta
     1321 gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     1381 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     1441 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     1501 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     1561 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc
     1621 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     1681 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     1741 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     1801 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     1861 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     1921 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     1981 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     2041 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggc tcaattagtc agcaaccata
     2101 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     2161 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     2221 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     2281 ggagcttgta tatccatttt cggaggttgg gcttcggaat cgttttccgg gacgccggct
     2341 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     2401 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     2461 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     2521 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     2581 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     2641 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     2701 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     2761 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     2821 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     2881 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     2941 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     3001 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     3061 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     3121 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     3181 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     3241 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     3301 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     3361 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     3421 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     3481 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     3541 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     3601 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     3661 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     3721 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     3781 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     3841 ccagtgctgc aatgataccg cgagatccac gctcaccggc tccagattta tcagcaataa
     3901 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     3961 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     4021 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     4081 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     4141 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     4201 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     4261 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     4321 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     4381 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     4441 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     4501 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     4561 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     4621 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     4681 ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
