LOCUS       sequence-318349-        6424 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression of intN-ZF10-NLS-HA under the EF1-alpha 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6424)
  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 6424)
  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: pJM589 EF1-alpha intN-ZF10-NLS-HA in TUPV2
FEATURES             Location/Qualifiers
     source          1..6424
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        202..1384
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          433..1375
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1332..1352
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     1386..1410
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2082..2102
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2109..2135
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2183..2200)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2189..2413
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3182..3610
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3269..3288)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3479..3500
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3633..3829
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3680..3815
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3742..3761
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    3949..4070
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3986..4005)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4040..4059
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4119..4135)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4119..4135)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4132..4154)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4143..4159
                     /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(4167..4173,4174..4191,4192..4197))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4212..4233
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4350..4367)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4521..5109)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4601..4620)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5280..6071,6072..6140))
                     /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     5903..5922
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6141..6245)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtg agtaattcat
       61 acaaaaggac tcgcccctgc cttggggaat cccagggacc gtcgttaaac tcccactaac
      121 gtagaaccca gagatcgctg cgttcccgcc ccctcacccg cccgctctcg tcatcactga
      181 ggtggagaag agcatgcgtg aggctccggt gcccgtcagt gggcagagcg cacatcgccc
      241 acagtccccg agaagttggg gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg
      301 cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg
      361 ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc
      421 gccagaacac aggtaagtgc cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat
      481 ggcccttgcg tgccttgaat tacttccacg cccctggctg cagtacgtga ttcttgatcc
      541 cgagcttcgg gttggaagtg ggtgggagag ttcgaggcct tgcgcttaag gagccccttc
      601 gcctcgtgct tgagttgagg cctggcttgg gcgctggggc cgccgcgtgc gaatctggtg
      661 gcaccttcgc gcctgtctcg ctgctttcga taagtctcta gccatttaaa atttttgatg
      721 acctgctgcg acgctttttt tctggcaaga tagtcttgta aatgcgggcc aagatctgca
      781 cactggtatt tcggtttttg gggccgcggg cggcgacggg gcccgtgcgt cccagcgcac
      841 atgttcggcg aggcggggcc tgcgagcgcg gccaccgaga atcggacggg ggtagtctca
      901 agctggccgg cctgctctgg tgcctggcct cgcgccgccg tgtatcgccc cgccctgggc
      961 ggcaaggctg gcccggtcgg caccagttgc gtgagcggaa agatggccgc ttcccggccc
     1021 tgctgcaggg agctcaaaat ggaggacgcg gcgctcggga gagcgggcgg gtgagtcacc
     1081 cacacaaagg aaaagggcct ttccgtcctc agccgtcgct tcatgtgact ccacggagta
     1141 ccgggcgccg tccaggcacc tcgattagtt ctcgagcttt tggagtacgt cgtctttagg
     1201 ttggggggag gggttttatg cgatggagtt tccccacact gagtgggtgg agactgaagt
     1261 taggccagct tggcacttga tgtaattctc cttggaattt gccctttttg agtttggatc
     1321 ttggttcatt ctcaagcctc agacagtggt tcaaagtttt tttcttccat ttcaggtgtc
     1381 gtgagagctc gtttagtgaa ccgtcagatc gcctggagac gccatccacg ctgttttgac
     1441 ctccatagaa gagaacggga ccgatccaaa ttctcgaggc tagcgccacc atgaccagaa
     1501 gcggatactg cctggacctg aagacccagg tgcagacccc tcagggcatg aaggagatca
     1561 gcaacatcca ggtgggcgac ctggtgctga gcaacaccgg ctacaacgag gtgctgaacg
     1621 tgttccccaa gagcaagaag aagagctaca agatcaccct ggaggacggc aaggagatca
     1681 tctgcagcga ggagcacctg ttccccaccc agaccggcga gatgaacatc agcggcggcc
     1741 tgaaggaggg catgtgcctg tacgtgaagg agactggttc tggttctggt tcaggatccg
     1801 gtaccggtgg agctagaccc ggggagcgcc ccttccagtg tcgcatttgc atgcggaact
     1861 tttcgactaa tcaaaaattg gaagttcata cccgtactca taccggtgaa aaaccgtttc
     1921 agtgtcggat ctgtatgcga aatttctccg ttagacataa tttgcaaaga catctacgta
     1981 cgcacaccgg cgagaagcca ttccaatgcc gaatatgcat gcgcaacttc agtcaacatc
     2041 caaatttgac tagacaccta aaaacccacc tgagaggatc tcctaagaag aagcgcaaag
     2101 ttggcatgta cccatacgac gtaccagatt acgcttaggc ggccgctcga gtctagaggg
     2161 cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     2221 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     2281 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     2341 gtggggcagg acagcaaggg ggaggattgg gaagtcaata gcaggcatgc tggggatgcg
     2401 gtgggctcta tggcttctga ggcaggtcga agaactattt cctggctgtt acgcgagcgc
     2461 atcgcgataa gctctagagg gacagccccc ccccaaagcc cccagggatg taattacgtc
     2521 cctcccccgc tagggggcag cagcgagccg cccggggctc cgctccggtc cggcgctccc
     2581 cccgcatccc cgagccggca gcgtgcgggg acagcccggg cacggggaag gtggcacggg
     2641 atcgctttcc tctgaacgct tctcgctgct ctttgagcct gcagacacct ggggggatac
     2701 ggggaaaaag ctttaggctg aaagagagat ttagaatgac agaatcatag aacggcctgc
     2761 gtacggatgg ggctctagct ctagagcttg atatcgaatt cctgcagccc cgctagaggg
     2821 acagcccccc cccaaagccc ccagggatgt aattacgtcc ctcccccgct agggggcagc
     2881 agcgagccgc ccggggctcc gctccggtcc ggcgctcccc ccgcatcccc gagccggcag
     2941 cgtgcgggga cagcccgggc acggggaagg tggcacggga tcgctttcct ctgaacgctt
     3001 ctcgctgctc tttgagcctg cagacacctg gggggatacg gggaaaaagc tttaggctga
     3061 aagagagatt tagaatgaca gaatcataga acggcctggg cgcgcctggt ggcgaacgat
     3121 caacagacgg gagaggacta aagtcttcgc tgacagcgac cggactctag ggggtatccc
     3181 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     3241 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     3301 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     3361 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     3421 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     3481 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     3541 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     3601 aacgcgaatt aattctgtgg aatgtgtgtc agttaggctc aattagtcag caaccatagt
     3661 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     3721 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
     3781 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
     3841 agcttgtata tccattttcg gaggttgggc ttcggaatcg ttttccggga cgccggctgg
     3901 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     3961 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     4021 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     4081 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     4141 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     4201 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     4261 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     4321 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     4381 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     4441 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     4501 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     4561 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     4621 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     4681 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     4741 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     4801 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     4861 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     4921 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     4981 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     5041 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     5101 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     5161 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     5221 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     5281 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     5341 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     5401 agtgctgcaa tgataccgcg agatccacgc tcaccggctc cagatttatc agcaataaac
     5461 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     5521 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     5581 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     5641 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     5701 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     5761 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     5821 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     5881 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     5941 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     6001 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     6061 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     6121 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     6181 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     6241 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tcgaagacgg
     6301 gcaaaaccga tgttgacgga ctaatcctga caagatctgg ggaggtcgct gagtagtgcg
     6361 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
     6421 ttag
//
