LOCUS       sequence-323915-        4662 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EYFP reporter vector for the ZF43x10-Spaced-Far promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4662)
  TITLE     Leonard Lab COMET plasmids
REFERENCE   2  (bases 1 to 4662)
  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: pPD148 ZF43x10-Spaced-Far_EYFP
FEATURES             Location/Qualifiers
     source          1..4662
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      522..531
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(528..530,531..533,534..1247)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(573..594)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(834..853)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1181..1202
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1292..1309)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1298..1522
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1568..1996
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1655..1674)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1865..1886
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2019..2215
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2066..2201
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2128..2147
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    2335..2456
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2372..2391)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2426..2445
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2505..2521)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2505..2521)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2518..2540)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2529..2545
                     /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(2553..2559,2560..2577,2578..2583))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2598..2619
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2736..2753)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2907..3495)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2987..3006)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3666..4457,4458..4526))
                     /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     4289..4308
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4527..4631)
                     /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 gatccctcta gagggagatc ctgagtagtg cgcgagcaaa
      361 atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt
      421 aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgatccctct agagggtata
      481 taatgggggc cactagtcta ctaccagagc tcatcgctag cgccaccatg gtgagcaagg
      541 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
      601 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
      661 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccacct
      721 tcggctacgg cctgcagtgc ttcgcccgct accccgacca catgaagcag cacgacttct
      781 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
      841 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
      901 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
      961 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga
     1021 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
     1081 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagctacc
     1141 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     1201 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaggcg gccgctcgag
     1261 tctagagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
     1321 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     1381 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     1441 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     1501 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     1561 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     1621 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     1681 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     1741 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     1801 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     1861 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     1921 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     1981 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt aggctcaatt agtcagcaac
     2041 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     2101 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     2161 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     2221 cccgggagct tgtatatcca ttttcggagg ttgggcttcg gaatcgtttt ccgggacgcc
     2281 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     2341 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     2401 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     2461 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     2521 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     2581 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     2641 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     2701 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     2761 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     2821 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     2881 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     2941 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     3001 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     3061 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     3121 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     3181 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     3241 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     3301 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     3361 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     3421 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     3481 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     3541 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     3601 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     3661 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     3721 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     3781 ggccccagtg ctgcaatgat accgcgagat ccacgctcac cggctccaga tttatcagca
     3841 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     3901 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     3961 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     4021 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     4081 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     4141 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     4201 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     4261 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     4321 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     4381 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     4441 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     4501 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     4561 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     4621 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
