LOCUS       sequence-324231-        4487 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EYFP reporter vector for the ZF43x16-Compact promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4487)
  TITLE     Leonard Lab COMET plasmids
REFERENCE   2  (bases 1 to 4487)
  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: pPD301 ZF43x16-Compact_EYFP
FEATURES             Location/Qualifiers
     source          1..4487
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      362..371
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(368..370,371..373,374..1087)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(413..434)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(674..693)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1021..1042
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1132..1149)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1138..1362
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1408..1836
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1495..1514)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1705..1726
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1859..2055
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1906..2041
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1968..1987
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    2175..2296
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2212..2231)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2266..2285
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2345..2361)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2345..2361)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2358..2380)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2369..2385
                     /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(2393..2399,2400..2417,2418..2423))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2438..2459
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2576..2593)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2747..3335)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2827..2846)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3506..4297,4298..4366))
                     /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     4129..4148
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4367..4471)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtgccacctg acgtcgacgg atcgggagat ctgaatttga gaccacagtg atacggtctc
       61 cgatcggagt gaggaaaatt agagtgagga cgatcggagt gaggataatt agagtgagga
      121 cgatcggagt gaggaaaatt agagtgagga cgatcggagt gaggataatt agagtgagga
      181 cgatcggagt gaggaaaatt agagtgagga cgatcggagt gaggataatt agagtgagga
      241 cgatcggagt gaggaaaatt agagtgagga cgatcggagt gaggataatt agagtgagga
      301 cgatccctct agagggtata taatgggggc cactagtcta ctaccagagc tcatcgctag
      361 cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
      421 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
      481 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
      541 gcccaccctc gtgaccacct tcggctacgg cctgcagtgc ttcgcccgct accccgacca
      601 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
      661 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
      721 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
      781 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
      841 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
      901 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
      961 caaccactac ctgagctacc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     1021 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     1081 caagtaggcg gccgctcgag tctagagggc ccgtttaaac ccgctgatca gcctcgactg
     1141 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     1201 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     1261 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     1321 aagtcaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
     1381 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
     1441 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     1501 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     1561 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     1621 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     1681 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     1741 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     1801 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
     1861 aggctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     1921 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     1981 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     2041 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggagg ttgggcttcg
     2101 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     2161 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     2221 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     2281 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     2341 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     2401 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     2461 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     2521 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     2581 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     2641 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     2701 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     2761 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     2821 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     2881 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     2941 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     3001 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     3061 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     3121 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     3181 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     3241 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     3301 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     3361 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     3421 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     3481 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     3541 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     3601 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagat ccacgctcac
     3661 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     3721 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     3781 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     3841 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     3901 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     3961 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     4021 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     4081 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     4141 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     4201 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     4261 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     4321 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     4381 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     4441 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaa
//
