LOCUS       sequence-318785-        5763 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of 3xFLAG-NLS-DsDed-ZF1-PEST under the ZF10x6-C
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5763)
  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 5763)
  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: pJM553 ZF10x6-C 3xFLAG-NLS-DsDed-ZF1-PEST in T...
FEATURES             Location/Qualifiers
     source          1..5763
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      357..366
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             366..431
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             438..458
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             501..1175
                     /codon_start=1
                     /product="noncytotoxic tetrameric variant of DsRed 
                     fluorescent protein (Strack et al., 2008)"
                     /label=DsRed-Express2
                     /note="mammalian codon-optimized"
                     /translation="MDSTENVIKPFMRFKVHMEGSVNGHEFEIEGEGEGKPYEGTQTAK
                     LQVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWEKVMNFEDGGV
                     VTVTQDSSLQDGTFIYHVKFIGVNFPSDGPVMQKKTLGWEPSTERLYPRDGVLKGEIHK
                     ALKLKGGGHYLVEFKSIYMAKKPVKLPGYYYVDSKLDITSHNEDYTVVEQYERAEARHH
                     LFQ"
     primer_bind     complement(681..701)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     1093..1116
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(1670..1687)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1676..1900
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2669..3097
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2756..2775)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2966..2987
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3120..3316
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3167..3302
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3229..3248
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    3436..3557
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3473..3492)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3527..3546
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3606..3622)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3606..3622)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3619..3641)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3630..3646
                     /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(3654..3660,3661..3678,3679..3684))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3699..3720
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3837..3854)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4008..4596)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4088..4107)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4767..5558,5559..5627))
                     /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     5390..5409
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5628..5732)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatcgaag acgggcaaaa ccgatgttga cggactaatc ctgacaagat
       61 ctggggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
      121 ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac
      181 gggccagata tacgcgtggg agatctgcga ggaagatggt acgcgagaag atggttgagc
      241 tgaagatggt gactaagaag atggtacgcg agaagatggt tgagctgaag atggtaacgc
      301 tctctagagg gtatataatg ggggccacta gtctactacc agagctcatc gctagcgcca
      361 ccatggatta taaggatcac gatggcgact acaaggacca cgatattgac tacaaagacg
      421 atgacgataa aatggctcct aagaaaaagc gcaaagtcgg tatccatggc gttcccggat
      481 caggctccgg aagtggctcc atggatagca ctgagaacgt catcaagccc ttcatgcgct
      541 tcaaggtgca catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg
      601 agggcaagcc ctacgagggc acccagaccg ccaagctcca ggtgaccaag ggcggccccc
      661 tgcccttcgc ctgggacatc ctgtcccccc agttccagta cggctccaag gtgtacgtga
      721 agcaccccgc cgacatcccc gactacaaga agctgtcctt ccccgagggc ttcaagtggg
      781 agaaggtgat gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctcc
      841 aggacggcac cttcatctac cacgtgaagt tcatcggcgt gaacttcccc tccgacggcc
      901 ccgtaatgca gaagaaaact ctgggctggg agccctccac cgagcgcctg tacccccgcg
      961 acggcgtgct gaagggcgag atccacaagg cgctgaagct gaagggcggc ggccactacc
     1021 tggtggagtt caagtcaatc tacatggcca agaagcccgt gaagctgccc ggctactact
     1081 acgtggactc caagctggac atcacctccc acaacgagga ctacaccgtg gtggagcagt
     1141 acgagcgcgc cgaggcccgc caccacctgt tccagtctgg ttctggttct ggttcaggat
     1201 ccggtaccgg tggagctaga cccggggagc gccccttcca gtgtcgcatt tgcatgcgga
     1261 acttttcgcg ccaggacagg cttgacaggc atacccgtac tcataccggt gaaaaaccgt
     1321 ttcagtgtcg gatctgtatg cgaaatttct cccagaagga gcacttggcg gggcatctac
     1381 gtacgcacac cggcgagaag ccattccaat gccgaatatg catgcgcaac ttcagtcgcc
     1441 gcgacaacct gaaccggcac ctaaaaaccc acctgagagg atctggttct ggatcaggta
     1501 gtagtcatgg atttccacct gaagtcgagg aacaggatga tggcaccctc ccaatgagct
     1561 gtgcccaaga gtccgggatg gaccgccatc cagcagcttg cgcatcagct cgcataaatg
     1621 tttaggcggc cgctcgagtc tagagggccc gtttaaaccc gctgatcagc ctcgactgtg
     1681 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     1741 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     1801 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     1861 gtcaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc aggtcgaaga
     1921 actatttcct ggctgttacg cgagcgcatc gcgataagct ctagagggac agcccccccc
     1981 caaagccccc agggatgtaa ttacgtccct cccccgctag ggggcagcag cgagccgccc
     2041 ggggctccgc tccggtccgg cgctcccccc gcatccccga gccggcagcg tgcggggaca
     2101 gcccgggcac ggggaaggtg gcacgggatc gctttcctct gaacgcttct cgctgctctt
     2161 tgagcctgca gacacctggg gggatacggg gaaaaagctt taggctgaaa gagagattta
     2221 gaatgacaga atcatagaac ggcctgcgta cggatggggc tctagctcta gagcttgata
     2281 tcgaattcct gcagccccgc tagagggaca gccccccccc aaagccccca gggatgtaat
     2341 tacgtccctc ccccgctagg gggcagcagc gagccgcccg gggctccgct ccggtccggc
     2401 gctccccccg catccccgag ccggcagcgt gcggggacag cccgggcacg gggaaggtgg
     2461 cacgggatcg ctttcctctg aacgcttctc gctgctcttt gagcctgcag acacctgggg
     2521 ggatacgggg aaaaagcttt aggctgaaag agagatttag aatgacagaa tcatagaacg
     2581 gcctgggcgc gcctggtggc gaacgatcaa cagacgggag aggactaaag tcttcgctga
     2641 cagcgaccgg actctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     2701 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     2761 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     2821 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     2881 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     2941 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     3001 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     3061 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     3121 taggctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     3181 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     3241 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     3301 cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggag gttgggcttc
     3361 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     3421 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     3481 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     3541 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     3601 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     3661 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     3721 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     3781 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     3841 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     3901 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     3961 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     4021 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     4081 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     4141 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     4201 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     4261 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     4321 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     4381 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     4441 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     4501 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     4561 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     4621 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     4681 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     4741 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     4801 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     4861 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga tccacgctca
     4921 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     4981 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     5041 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     5101 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     5161 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     5221 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     5281 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     5341 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     5401 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     5461 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     5521 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     5581 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     5641 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     5701 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     5761 gtc
//
