LOCUS       sequence-323881-        4606 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EYFP reporter vector for the ZF43x8-Spaced-Far promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4606)
  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 4606)
  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: pPD147 ZF43x8-Spaced-Far_EYFP
FEATURES             Location/Qualifiers
     source          1..4606
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      466..475
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(472..474,475..477,478..1191)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(517..538)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(778..797)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1125..1146
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1236..1253)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1242..1466
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1512..1940
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1599..1618)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1809..1830
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1963..2159
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2010..2145
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2072..2091
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    2279..2400
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2316..2335)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2370..2389
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2449..2465)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2449..2465)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2462..2484)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2473..2489
                     /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(2497..2503,2504..2521,2522..2527))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2542..2563
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2680..2697)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2851..3439)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2931..2950)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3610..4401,4402..4470))
                     /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     4233..4252
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4471..4575)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctgaa ttcggatcca gagtgaggac tacctataga agtaagagtg
       61 aggacaatcg tatacaagta gatccagagt gaggactacc tatagaagta agagtgagga
      121 caatcgtata caagtagatc cctctagagg gagatccaga gtgaggacta cctatagaag
      181 taagagtgag gacaatcgta tacaagtaga tccagagtga ggactaccta tagaagtaag
      241 agtgaggaca atcgtataca agtagatccc tctagaggga gatcctgagt agtgcgcgag
      301 caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag
      361 ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgatcc ctctagaggg
      421 tatataatgg gggccactag tctactacca gagctcatcg ctagcgccac catggtgagc
      481 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
      541 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
      601 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
      661 accttcggct acggcctgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac
      721 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
      781 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
      841 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
      901 tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag
      961 gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac
     1021 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     1081 taccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     1141 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta ggcggccgct
     1201 cgagtctaga gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     1261 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     1321 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     1381 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     1441 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     1501 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     1561 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     1621 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     1681 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     1741 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     1801 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     1861 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     1921 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttaggctc aattagtcag
     1981 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     2041 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     2101 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     2161 agctcccggg agcttgtata tccattttcg gaggttgggc ttcggaatcg ttttccggga
     2221 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     2281 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     2341 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     2401 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     2461 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     2521 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     2581 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     2641 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     2701 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     2761 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     2821 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     2881 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     2941 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3001 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     3061 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3121 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3181 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3241 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     3301 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     3361 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     3421 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     3481 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     3541 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     3601 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     3661 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     3721 atctggcccc agtgctgcaa tgataccgcg agatccacgc tcaccggctc cagatttatc
     3781 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     3841 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     3901 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     3961 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     4021 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     4081 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     4141 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     4201 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     4261 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     4321 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     4381 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     4441 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     4501 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     4561 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
