LOCUS       sequence-317641-        5481 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of 3xFLAG-NLS-DsDed-ZF1 under the ZF2x6-C 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5481)
  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 5481)
  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: pJM574 ZF2x6-C 3xFLAG-NLS-DsDed-ZF1 in TUPV2
FEATURES             Location/Qualifiers
     source          1..5481
                     /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             253..318
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             325..345
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             388..1062
                     /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(568..588)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     980..1003
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(1419..1436)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1425..1649
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2418..2846
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2505..2524)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2715..2736
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2869..3065
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2916..3051
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2978..2997
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    3185..3306
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3222..3241)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3276..3295
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3355..3371)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3355..3371)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3368..3390)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3379..3395
                     /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(3403..3409,3410..3427,3428..3433))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3448..3469
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3586..3603)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3757..4345)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3837..3856)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4516..5307,5308..5376))
                     /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     5139..5158
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5377..5481)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatcgaa gacgggcaaa
       61 accgatgttg acggactaat cctgacaaga tctgcgagga ggacgtgacg cgagaggacg
      121 tgtgagctga ggacgtggac taagaggacg tgacgcgaga ggacgtgtga gctgaggacg
      181 tgaacgctct ctagagggta tataatgggg gccactagtc tactaccaga gctcatcgct
      241 agcgccacca tggattataa ggatcacgat ggcgactaca aggaccacga tattgactac
      301 aaagacgatg acgataaaat ggctcctaag aaaaagcgca aagtcggtat ccatggcgtt
      361 cccggatcag gctccggaag tggctccatg gatagcactg agaacgtcat caagcccttc
      421 atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
      481 gagggcgagg gcaagcccta cgagggcacc cagaccgcca agctccaggt gaccaagggc
      541 ggccccctgc ccttcgcctg ggacatcctg tccccccagt tccagtacgg ctccaaggtg
      601 tacgtgaagc accccgccga catccccgac tacaagaagc tgtccttccc cgagggcttc
      661 aagtgggaga aggtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc
      721 tccctccagg acggcacctt catctaccac gtgaagttca tcggcgtgaa cttcccctcc
      781 gacggccccg taatgcagaa gaaaactctg ggctgggagc cctccaccga gcgcctgtac
      841 ccccgcgacg gcgtgctgaa gggcgagatc cacaaggcgc tgaagctgaa gggcggcggc
      901 cactacctgg tggagttcaa gtcaatctac atggccaaga agcccgtgaa gctgcccggc
      961 tactactacg tggactccaa gctggacatc acctcccaca acgaggacta caccgtggtg
     1021 gagcagtacg agcgcgccga ggcccgccac cacctgttcc agtctggttc tggttctggt
     1081 tcaggatccg gtaccggtgg agctagaccc ggggagcgcc ccttccagtg tcgcatttgc
     1141 atgcggaact tttcgcgcca ggacaggctt gacaggcata cccgtactca taccggtgaa
     1201 aaaccgtttc agtgtcggat ctgtatgcga aatttctccc agaaggagca cttggcgggg
     1261 catctacgta cgcacaccgg cgagaagcca ttccaatgcc gaatatgcat gcgcaacttc
     1321 agtcgccgcg acaacctgaa ccggcaccta aaaacccacc tgagaggatc ttaagcggcc
     1381 gctcgagtct agagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg
     1441 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     1501 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     1561 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag tcaatagcag
     1621 gcatgctggg gatgcggtgg gctctatggc ttctgaggca ggtcgaagaa ctatttcctg
     1681 gctgttacgc gagcgcatcg cgataagctc tagagggaca gccccccccc aaagccccca
     1741 gggatgtaat tacgtccctc ccccgctagg gggcagcagc gagccgcccg gggctccgct
     1801 ccggtccggc gctccccccg catccccgag ccggcagcgt gcggggacag cccgggcacg
     1861 gggaaggtgg cacgggatcg ctttcctctg aacgcttctc gctgctcttt gagcctgcag
     1921 acacctgggg ggatacgggg aaaaagcttt aggctgaaag agagatttag aatgacagaa
     1981 tcatagaacg gcctgcgtac ggatggggct ctagctctag agcttgatat cgaattcctg
     2041 cagccccgct agagggacag ccccccccca aagcccccag ggatgtaatt acgtccctcc
     2101 cccgctaggg ggcagcagcg agccgcccgg ggctccgctc cggtccggcg ctccccccgc
     2161 atccccgagc cggcagcgtg cggggacagc ccgggcacgg ggaaggtggc acgggatcgc
     2221 tttcctctga acgcttctcg ctgctctttg agcctgcaga cacctggggg gatacgggga
     2281 aaaagcttta ggctgaaaga gagatttaga atgacagaat catagaacgg cctgggcgcg
     2341 cctggtggcg aacgatcaac agacgggaga ggactaaagt cttcgctgac agcgaccgga
     2401 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     2461 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     2521 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     2581 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     2641 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     2701 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     2761 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     2821 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt aggctcaatt
     2881 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     2941 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     3001 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     3061 gcaaaaagct cccgggagct tgtatatcca ttttcggagg ttgggcttcg gaatcgtttt
     3121 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     3181 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     3241 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     3301 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     3361 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     3421 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     3481 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     3541 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     3601 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     3661 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     3721 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3781 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3841 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     3901 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     3961 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4021 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4081 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4141 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     4201 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4261 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     4321 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     4381 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     4441 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     4501 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     4561 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     4621 cttaccatct ggccccagtg ctgcaatgat accgcgagat ccacgctcac cggctccaga
     4681 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     4741 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     4801 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     4861 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     4921 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     4981 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     5041 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     5101 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     5161 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     5221 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     5281 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     5341 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     5401 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     5461 taaacaaata ggggttccgc g
//
