LOCUS       sequence-317929-        6289 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression of intC-ZF1-NLS-HA under the EF1-alpha 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6289)
  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 6289)
  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: pJM516 EF1-alpha intC-ZF1-NLS-HA in TUPV2
FEATURES             Location/Qualifiers
     source          1..6289
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          231..1173
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1130..1150
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     1184..1208
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1739..1759
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1772..1798
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(1846..1863)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1852..2076
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2845..3273
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2932..2951)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3142..3163
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3296..3492
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3343..3478
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3405..3424
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    3612..3733
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3649..3668)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3703..3722
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3782..3798)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3782..3798)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3795..3817)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3806..3822
                     /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(3830..3836,3837..3854,3855..3860))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3875..3896
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4013..4030)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4184..4772)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4264..4283)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4943..5734,5735..5803))
                     /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     5566..5585
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5804..5908)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(6289,1..1182)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
ORIGIN
        1 gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg
       61 gaggggtcgg caattgaacc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg
      121 atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag
      181 tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg
      241 tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg ccttgaatta
      301 cttccacgcc cctggctgca gtacgtgatt cttgatcccg agcttcgggt tggaagtggg
      361 tgggagagtt cgaggccttg cgcttaagga gccccttcgc ctcgtgcttg agttgaggcc
      421 tggcttgggc gctggggccg ccgcgtgcga atctggtggc accttcgcgc ctgtctcgct
      481 gctttcgata agtctctagc catttaaaat ttttgatgac ctgctgcgac gctttttttc
      541 tggcaagata gtcttgtaaa tgcgggccaa gatctgcaca ctggtatttc ggtttttggg
      601 gccgcgggcg gcgacggggc ccgtgcgtcc cagcgcacat gttcggcgag gcggggcctg
      661 cgagcgcggc caccgagaat cggacggggg tagtctcaag ctggccggcc tgctctggtg
      721 cctggcctcg cgccgccgtg tatcgccccg ccctgggcgg caaggctggc ccggtcggca
      781 ccagttgcgt gagcggaaag atggccgctt cccggccctg ctgcagggag ctcaaaatgg
      841 aggacgcggc gctcgggaga gcgggcgggt gagtcaccca cacaaaggaa aagggccttt
      901 ccgtcctcag ccgtcgcttc atgtgactcc acggagtacc gggcgccgtc caggcacctc
      961 gattagttct cgagcttttg gagtacgtcg tctttaggtt ggggggaggg gttttatgcg
     1021 atggagtttc cccacactga gtgggtggag actgaagtta ggccagcttg gcacttgatg
     1081 taattctcct tggaatttgc cctttttgag tttggatctt ggttcattct caagcctcag
     1141 acagtggttc aaagtttttt tcttccattt caggtgtcgt gagagctcgt ttagtgaacc
     1201 gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga gaacgggacc
     1261 gatccaaatt ctcgaggcta gcgccaccat gatgctgaag aagatcctga agatcgagga
     1321 gctggacgag agagagctga tcgacatcga ggtgagcggc aaccacctgt tctacgccaa
     1381 cgacatcctg acccacaaca gcggatccgg tggtggcggg tctggaggag gaggtagtgg
     1441 tggaggtggc tctggtaccg ctagacccgg ggagcgcccc ttccagtgtc gcatttgcat
     1501 gcggaacttt tcgcgccagg acaggcttga caggcatacc cgtactcata ccggtgaaaa
     1561 accgtttcag tgtcggatct gtatgcgaaa tttctcccag aaggagcact tggcggggca
     1621 tctacgtacg cacaccggcg agaagccatt ccaatgccga atatgcatgc gcaacttcag
     1681 tcgccgcgac aacctgaacc ggcacctaaa aacccacctg agagaattcg gctctggccc
     1741 taagaagaag cgcaaagttg gctctggcat gtacccatac gacgtaccag attacgctta
     1801 ggcggccgct cgagtctaga gggcccgttt aaacccgctg atcagcctcg actgtgcctt
     1861 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     1921 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     1981 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagtca
     2041 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcaggt cgaagaacta
     2101 tttcctggct gttacgcgag cgcatcgcga taagctctag agggacagcc cccccccaaa
     2161 gcccccaggg atgtaattac gtccctcccc cgctaggggg cagcagcgag ccgcccgggg
     2221 ctccgctccg gtccggcgct ccccccgcat ccccgagccg gcagcgtgcg gggacagccc
     2281 gggcacgggg aaggtggcac gggatcgctt tcctctgaac gcttctcgct gctctttgag
     2341 cctgcagaca cctgggggga tacggggaaa aagctttagg ctgaaagaga gatttagaat
     2401 gacagaatca tagaacggcc tgcgtacgga tggggctcta gctctagagc ttgatatcga
     2461 attcctgcag ccccgctaga gggacagccc ccccccaaag cccccaggga tgtaattacg
     2521 tccctccccc gctagggggc agcagcgagc cgcccggggc tccgctccgg tccggcgctc
     2581 cccccgcatc cccgagccgg cagcgtgcgg ggacagcccg ggcacgggga aggtggcacg
     2641 ggatcgcttt cctctgaacg cttctcgctg ctctttgagc ctgcagacac ctggggggat
     2701 acggggaaaa agctttaggc tgaaagagag atttagaatg acagaatcat agaacggcct
     2761 gggcgcgcct ggtggcgaac gatcaacaga cgggagagga ctaaagtctt cgctgacagc
     2821 gaccggactc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     2881 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     2941 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     3001 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     3061 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     3121 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     3181 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     3241 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg
     3301 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     3361 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     3421 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     3481 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggaggttg ggcttcggaa
     3541 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     3601 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     3661 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     3721 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     3781 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     3841 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     3901 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     3961 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     4021 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     4081 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     4141 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     4201 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     4261 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     4321 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     4381 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     4441 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     4501 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     4561 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     4621 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     4681 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     4741 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     4801 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     4861 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     4921 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     4981 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     5041 gggagggctt accatctggc cccagtgctg caatgatacc gcgagatcca cgctcaccgg
     5101 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     5161 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     5221 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     5281 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     5341 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     5401 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     5461 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     5521 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     5581 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     5641 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     5701 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     5761 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     5821 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     5881 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg
     5941 acggatcggg agatcgaaga cgggcaaaac cgatgttgac ggactaatcc tgacaagatc
     6001 tggggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac
     6061 cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg
     6121 ggccagatat acgcgtgagt aattcataca aaaggactcg cccctgcctt ggggaatccc
     6181 agggaccgtc gttaaactcc cactaacgta gaacccagag atcgctgcgt tcccgccccc
     6241 tcacccgccc gctctcgtca tcactgaggt ggagaagagc atgcgtgag
//
