LOCUS       sequence-318439-        5107 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of EYFP under the ZF1x6-C promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5107)
  AUTHORS   Muldoon JJ, Kandula V, Hong M, Donahue PS, Boucher JD, Bagheri N, 
            Leonard JN
  TITLE     Model-guided design of mammalian genetic programs.
  JOURNAL   Sci Adv. 2021 Feb 19;7(8). pii: 7/8/eabe9375. doi: 
            10.1126/sciadv.abe9375. Print 2021 Feb.
  PUBMED    33608279
REFERENCE   2  (bases 1 to 5107)
  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: "Sci Adv."; 
            date: "2021-02-19"; volume: "7(8). pii: 7/8/eabe9375. doi"; pages: "
            10.1126/sciadv.abe9375. Print 2021 Feb"
COMMENT     Sequence Label: pJM502 ZF1x6-C EYFP in TUPV1
FEATURES             Location/Qualifiers
     source          1..5107
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      244..253
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(250..252,253..255,256..969)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(295..316)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(556..575)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     903..924
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1014..1031)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1020..1244
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2013..2441
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2100..2119)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2310..2331
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2464..2660
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2511..2646
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2573..2592
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    2780..2901
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2817..2836)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2871..2890
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2950..2966)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2950..2966)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2963..2985)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2974..2990
                     /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(2998..3004,3005..3022,3023..3028))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3043..3064
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3181..3198)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3352..3940)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3432..3451)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4111..4902,4903..4971))
                     /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     4734..4753
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4972..5076)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatcgaag acggtgccaa ggtcttgaca aacgagtgct agtacaagat
       61 cttgacgtcg acggatcggg agatctgaat tacgatcgga gtgaggataa ttagagtgag
      121 gacgatcgga gtgaggaaaa ttagagtgag gacgatcgga gtgaggataa ttagagtgag
      181 gacgatccct ctagagggta tataatgggg gccactagtc tactaccaga gctcatcgct
      241 agcgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
      301 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
      361 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
      421 tggcccaccc tcgtgaccac cttcggctac ggcctgcagt gcttcgcccg ctaccccgac
      481 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
      541 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
      601 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
      661 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
      721 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
      781 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
      841 gacaaccact acctgagcta ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
      901 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
      961 tacaagtagg cggccgctcg agtctagagg gcccgtttaa acccgctgat cagcctcgac
     1021 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     1081 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     1141 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     1201 ggaagtcaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcaggtcg
     1261 agggtacatg cgccttactc cttgtgagcg catcgcgata agctctagag ggacagcccc
     1321 cccccaaagc ccccagggat gtaattacgt ccctcccccg ctagggggca gcagcgagcc
     1381 gcccggggct ccgctccggt ccggcgctcc ccccgcatcc ccgagccggc agcgtgcggg
     1441 gacagcccgg gcacggggaa ggtggcacgg gatcgctttc ctctgaacgc ttctcgctgc
     1501 tctttgagcc tgcagacacc tggggggata cggggaaaaa gctttaggct gaaagagaga
     1561 tttagaatga cagaatcata gaacggcctg cgtacggatg gggctctagc tctagagctt
     1621 gatatcgaat tcctgcagcc ccgctagagg gacagccccc ccccaaagcc cccagggatg
     1681 taattacgtc cctcccccgc tagggggcag cagcgagccg cccggggctc cgctccggtc
     1741 cggcgctccc cccgcatccc cgagccggca gcgtgcgggg acagcccggg cacggggaag
     1801 gtggcacggg atcgctttcc tctgaacgct tctcgctgct ctttgagcct gcagacacct
     1861 ggggggatac ggggaaaaag ctttaggctg aaagagagat ttagaatgac agaatcatag
     1921 aacggcctgg gcgcgcctgg tggcgaacga tcaacagacg ggagagggca aaagtcttcg
     1981 ctgacagcga ccggactcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     2041 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     2101 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     2161 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     2221 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     2281 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     2341 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     2401 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     2461 cagttaggct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     2521 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     2581 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     2641 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggaggttggg
     2701 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     2761 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     2821 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     2881 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     2941 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     3001 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     3061 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     3121 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     3181 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     3241 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     3301 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     3361 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     3421 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     3481 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     3541 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     3601 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     3661 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     3721 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     3781 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     3841 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     3901 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     3961 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     4021 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     4081 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     4141 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     4201 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagatccacg
     4261 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     4321 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     4381 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     4441 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     4501 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     4561 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     4621 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     4681 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     4741 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     4801 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     4861 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     4921 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     4981 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     5041 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     5101 tgacgtc
//
