LOCUS       sequence-318367-        6343 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Constitutive expression of 3xFLAG-NLS-ZF2-ABI1 under the CMV 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6343)
  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 6343)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pKD011 CMV 3xFLAG-NLS-ZF2-ABI1 in TUPV2
FEATURES             Location/Qualifiers
     source          1..6343
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     249..268
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        249..267
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      287..296
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             296..361
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             368..388
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(1672..1689)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1678..1902
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2671..3099
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2758..2777)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2968..2989
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3122..3318
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3169..3304
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3231..3250
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    3438..3559
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3475..3494)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3529..3548
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3608..3624)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3608..3624)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3621..3643)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3632..3648
                     /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(3656..3662,3663..3680,3681..3686))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3701..3722
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3839..3856)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4010..4598)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4090..4109)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4769..5560,5561..5629))
                     /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     5392..5411
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5630..5734)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        5964..6343
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctctctgg ctaactagag aacccactgc ttactggctt
      241 atcgaaatta atacgactca ctatagggag tcccaagctg gctagcgcca ccatggacta
      301 caaagaccac gacggcgatt acaaggatca tgacatcgat tacaaagacg atgacgataa
      361 gatggctcct aagaaaaagc gcaaagtctc cggagctaga cccggggagc gccccttcca
      421 gtgtcgcatt tgcatgcgga acttttcgag aaatttcatt ttgcaaagac atacccgtac
      481 tcataccggt gaaaaaccgt ttcagtgtcg gatctgtatg cgaaatttct ccgatagagc
      541 taatttgagg aggcatctgc gtacgcacac cggcgagaag ccattccaat gccgaatatg
      601 catgcgcaac ttcagtagac atgatcaatt gactagacac ctaaaaaccc acctgagagg
      661 atctggatcc ggtggtggcg ggtctggagg aggaggtagt ggtggaggtg gctctggtac
      721 catgacgcgg gtgcctttgt atggttttac ttcgatttgt ggaagaaggc ctgagatgga
      781 agctgctgtt tcgactatac caagattcct tcaatcttcc tctggttcga tgttagatgg
      841 tcggtttgat cctcaatccg ccgctcattt cttcggtgtt tacgacggcc atggcggttc
      901 tcaggtagcg aactattgta gagagaggat gcatttggct ttggcggagg agatagctaa
      961 ggagaaaccg atgctctgcg atggtgatac gtggctggag aagtggaaga aagctctttt
     1021 caactcgttc ctgagagttg actcggagat tgagtcagtt gcgccggaga cggttgggtc
     1081 aacgtcggtg gttgccgttg ttttcccgtc tcacatcttc gtcgctaact gcggtgactc
     1141 tagagccgtt ctttgccgcg gcaaaactgc acttccatta tccgttgacc ataaaccgga
     1201 tagagaagat gaagctgcga ggattgaagc cgcaggaggg aaagtgattc agtggaatgg
     1261 agctcgtgtt ttcggtgttc tcgccatgtc gagatccatt ggcgatagat acttgaaacc
     1321 atccatcatt cctgatcctg aagtgacggc tgtgaagaga gtaaaagaag atgattgtct
     1381 gattttggcg agtgacgggg tttgggatgt aatgacggat gaagaagcgt gtgagatggc
     1441 aaggaagcgg attctcttgt ggcacaagaa aaacgcggtg gctggggatg catcgttgct
     1501 cgcggatgag cggagaaagg aagggaaaga tcctgcggcg atgtccgcgg ctgagtattt
     1561 gtcaaagctg gcgatacaga gaggaagcaa agacaacata agtgtggtgg tggttgattt
     1621 gaagtaggcg gccgctcgag tctagagggc ccgtttaaac ccgctgatca gcctcgactg
     1681 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     1741 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     1801 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     1861 aagtcaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcaggtcgaa
     1921 gaactatttc ctggctgtta cgcgagcgca tcgcgataag ctctagaggg acagcccccc
     1981 cccaaagccc ccagggatgt aattacgtcc ctcccccgct agggggcagc agcgagccgc
     2041 ccggggctcc gctccggtcc ggcgctcccc ccgcatcccc gagccggcag cgtgcgggga
     2101 cagcccgggc acggggaagg tggcacggga tcgctttcct ctgaacgctt ctcgctgctc
     2161 tttgagcctg cagacacctg gggggatacg gggaaaaagc tttaggctga aagagagatt
     2221 tagaatgaca gaatcataga acggcctgcg tacggatggg gctctagctc tagagcttga
     2281 tatcgaattc ctgcagcccc gctagaggga cagccccccc ccaaagcccc cagggatgta
     2341 attacgtccc tcccccgcta gggggcagca gcgagccgcc cggggctccg ctccggtccg
     2401 gcgctccccc cgcatccccg agccggcagc gtgcggggac agcccgggca cggggaaggt
     2461 ggcacgggat cgctttcctc tgaacgcttc tcgctgctct ttgagcctgc agacacctgg
     2521 ggggatacgg ggaaaaagct ttaggctgaa agagagattt agaatgacag aatcatagaa
     2581 cggcctgggc gcgcctggtg gcgaacgatc aacagacggg agaggactaa agtcttcgct
     2641 gacagcgacc ggactctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     2701 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     2761 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     2821 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     2881 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     2941 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     3001 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     3061 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
     3121 gttaggctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     3181 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     3241 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     3301 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg aggttgggct
     3361 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     3421 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     3481 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     3541 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     3601 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     3661 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     3721 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     3781 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     3841 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     3901 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     3961 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4021 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4081 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4141 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4201 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4261 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4321 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4381 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4441 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     4501 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     4561 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     4621 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     4681 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     4741 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     4801 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     4861 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gatccacgct
     4921 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     4981 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     5041 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     5101 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     5161 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     5221 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     5281 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     5341 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     5401 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     5461 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     5521 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     5581 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     5641 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     5701 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     5761 acgtcgacgg atcgggagat cgaagacggg caaaaccgat gttgacggac taatcctgac
     5821 aagatctggg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg
     5881 cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga
     5941 tgtacgggcc agatatacgc gttgacattg attattgact agttattaat agtaatcaat
     6001 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
     6061 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
     6121 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
     6181 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
     6241 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
     6301 tacttggcag tacatctacg tattagtcat cgctattacc atg
//
