LOCUS       sequence-323879-        4534 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EYFP reporter vector for the ZF43x6-Spaced-Far promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4534)
  TITLE     Leonard Lab COMET plasmids
REFERENCE   2  (bases 1 to 4534)
  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: pPD146 ZF43x6-Spaced-Far_EYFP
FEATURES             Location/Qualifiers
     source          1..4534
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      394..403
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(400..402,403..405,406..1119)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(445..466)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(706..725)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1053..1074
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1164..1181)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1170..1394
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1440..1868
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1527..1546)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1737..1758
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1891..2087
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1938..2073
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2000..2019
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    2207..2328
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2244..2263)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2298..2317
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2377..2393)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2377..2393)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2390..2412)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2401..2417
                     /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(2425..2431,2432..2449,2450..2455))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2470..2491
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2608..2625)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2779..3367)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2859..2878)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3538..4329,4330..4398))
                     /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     4161..4180
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4399..4503)
                     /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 tcctgagtag tgcgcgagca aaatttaagc
      241 tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt
      301 tgcgctgctt cgcgatgtac gggccagata tacgatccct ctagagggta tataatgggg
      361 gccactagtc tactaccaga gctcatcgct agcgccacca tggtgagcaa gggcgaggag
      421 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
      481 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
      541 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cttcggctac
      601 ggcctgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
      661 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
      721 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
      781 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
      841 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
      901 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
      961 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta ccagtccgcc
     1021 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     1081 gccgggatca ctctcggcat ggacgagctg tacaagtagg cggccgctcg agtctagagg
     1141 gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     1201 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     1261 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     1321 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     1381 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     1441 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     1501 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     1561 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     1621 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     1681 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     1741 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     1801 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     1861 cgcgaattaa ttctgtggaa tgtgtgtcag ttaggctcaa ttagtcagca accatagtcc
     1921 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     1981 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     2041 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     2101 cttgtatatc cattttcgga ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     2161 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     2221 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     2281 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     2341 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     2401 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     2461 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     2521 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     2581 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     2641 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     2701 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     2761 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     2821 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     2881 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     2941 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3001 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3061 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3121 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3181 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     3241 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3301 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3361 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3421 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     3481 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     3541 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     3601 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     3661 tgctgcaatg ataccgcgag atccacgctc accggctcca gatttatcag caataaacca
     3721 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     3781 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     3841 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     3901 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     3961 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     4021 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     4081 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     4141 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     4201 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     4261 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     4321 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     4381 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     4441 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     4501 gcgcacattt ccccgaaaag tgccacctga cgtc
//
