LOCUS       sequence-317691-        5166 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of intC-ZF1-ZF2-NLS-HA under the ZF6x6-C 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5166)
  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 5166)
  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: pJM595 ZF6x6-C intC-ZF1-ZF2-NLS-HA in TUPV2
FEATURES             Location/Qualifiers
     source          1..5166
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..105)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1108..1128
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1135..1161
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(1209..1226)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1215..1439
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2208..2636
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2295..2314)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2505..2526
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2659..2855
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2706..2841
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2768..2787
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    2975..3096
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3012..3031)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3066..3085
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3145..3161)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3145..3161)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3158..3180)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3169..3185
                     /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(3193..3199,3200..3217,3218..3223))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3238..3259
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3376..3393)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3547..4135)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3627..3646)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4306..5097,5098..5166))
                     /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     4929..4948
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
       61 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      121 agtgccacct gacgtcgacg gatcgggaga tcgaagacgg gcaaaaccga tgttgacgga
      181 ctaatcctga caagatctgc gaggtgtagg ggacgcgagt gtaggggtga gctgtgtagg
      241 gggactaagt gtaggggacg cgagtgtagg ggtgagctgt gtaggggaac gctctctaga
      301 gggtatataa tgggggccac tagtctacta ccagagctca tcgctagcgc caccatgatg
      361 ctgaagaaga tcctgaagat cgaggagctg gacgagagag agctgatcga catcgaggtg
      421 agcggcaacc acctgttcta cgccaacgac atcctgaccc acaacagcgg atccggtggt
      481 ggcgggtctg gaggaggagg tagtggtgga ggtggctctg gtaccgctag acccggggag
      541 cgccccttcc agtgtcgcat ttgcatgcgg aacttttcgc gccaggacag gcttgacagg
      601 catacccgta ctcataccgg tgaaaaaccg tttcagtgtc ggatctgtat gcgaaatttc
      661 tcccagaagg agcacttggc ggggcatcta cgtacgcaca ccggcgagaa gccattccaa
      721 tgccgaatat gcatgcgcaa cttcagtcgc cgcgacaacc tgaaccggca cctaaaaacc
      781 cacctgagaa gttccggatc ttctggttct ggttctggtt caggatccgg taccggtgga
      841 gctagacccg gggagcgccc cttccagtgt cgcatttgca tgcggaactt ttcgagaaat
      901 ttcattttgc aaagacatac ccgtactcat accggtgaaa aaccgtttca gtgtcggatc
      961 tgtatgcgaa atttctccga tagagctaat ttgaggaggc atctgcgtac gcacaccggc
     1021 gagaagccat tccaatgccg aatatgcatg cgcaacttca gtagacatga tcaattgact
     1081 agacacctaa aaacccacct gagaggacct aagaagaagc gcaaagttgg catgtaccca
     1141 tacgacgtac cagattacgc ttaggcggcc gctcgagtct agagggcccg tttaaacccg
     1201 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     1261 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     1321 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     1381 caagggggag gattgggaag tcaatagcag gcatgctggg gatgcggtgg gctctatggc
     1441 ttctgaggca ggtcgaagaa ctatttcctg gctgttacgc gagcgcatcg cgataagctc
     1501 tagagggaca gccccccccc aaagccccca gggatgtaat tacgtccctc ccccgctagg
     1561 gggcagcagc gagccgcccg gggctccgct ccggtccggc gctccccccg catccccgag
     1621 ccggcagcgt gcggggacag cccgggcacg gggaaggtgg cacgggatcg ctttcctctg
     1681 aacgcttctc gctgctcttt gagcctgcag acacctgggg ggatacgggg aaaaagcttt
     1741 aggctgaaag agagatttag aatgacagaa tcatagaacg gcctgcgtac ggatggggct
     1801 ctagctctag agcttgatat cgaattcctg cagccccgct agagggacag ccccccccca
     1861 aagcccccag ggatgtaatt acgtccctcc cccgctaggg ggcagcagcg agccgcccgg
     1921 ggctccgctc cggtccggcg ctccccccgc atccccgagc cggcagcgtg cggggacagc
     1981 ccgggcacgg ggaaggtggc acgggatcgc tttcctctga acgcttctcg ctgctctttg
     2041 agcctgcaga cacctggggg gatacgggga aaaagcttta ggctgaaaga gagatttaga
     2101 atgacagaat catagaacgg cctgggcgcg cctggtggcg aacgatcaac agacgggaga
     2161 ggactaaagt cttcgctgac agcgaccgga ctctaggggg tatccccacg cgccctgtag
     2221 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     2281 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     2341 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     2401 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     2461 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     2521 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     2581 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     2641 ctgtggaatg tgtgtcagtt aggctcaatt agtcagcaac catagtcccg cccctaactc
     2701 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     2761 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     2821 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     2881 ttttcggagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     2941 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     3001 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     3061 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     3121 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     3181 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     3241 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     3301 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     3361 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     3421 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     3481 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     3541 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     3601 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     3661 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     3721 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     3781 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     3841 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     3901 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     3961 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     4021 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     4081 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     4141 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     4201 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     4261 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     4321 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     4381 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     4441 accgcgagat ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     4501 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     4561 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     4621 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     4681 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     4741 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     4801 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     4861 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     4921 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     4981 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     5041 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     5101 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     5161 actcat
//
