pNFAT-mGreenLantern-iPAK4

LOCUS       pNFAT-mGreenLantern-iPAK4            6142 bp DNA     circular SYN 05-DEC-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6142)
  AUTHORS   Rui
  TITLE     Direct Submission
  JOURNAL   Exported Jul 14, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..6142
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          958..2034
                     /mol_type="genomic DNA"
                     /organism="."
     misc_feature    20..37
                     /label=NFAT_IL4
     misc_feature    50..67
                     /label=NFAT_IL4
     misc_feature    80..97
                     /label=NFAT_IL4
     misc_feature    110..127
                     /label=NFAT_IL4
     misc_feature    140..157
                     /label=NFAT_IL4
     promoter        171..202
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     primer_bind     208..255
                     /label=NFAT-EcoRI-mGL-F
     regulatory      235..244
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             241..957
                     /codon_start=1
                     /label=mGreenLantern
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLGYGVACFARYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIVLKGIDFKEDGNILGHKLEYNFNSHKVYITADKQKNGIK
                     ANFKTRHNVEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLK
                     ERVTAAGITHDMDELYK"
     primer_bind     complement(942..983)
                     /label=INKA-mGreenLantern-R
     CDS             970..1083
                     /codon_start=1
                     /label=INKA-box
                     /translation="EAEDWTAALLNRGRSRQPLVLGDNCFADLVHNWMELPE"
     CDS             1090..2031
                     /codon_start=1
                     /label=Pak4cat
                     /translation="PAAPAVPGPPGPRSPQREPQRVSHEQFRAALQLVVDPGDPRSYLD
                     NFIKIGEGSTGIVCIATVRSSGKLVAVKKMDLRKQQRRELLFNEVVIMRDYQHENVVEM
                     YNSYLVGDELWVVMEFLEGGALTDIVTHTRMNEEQIAAVCLAVLQALSVLHAQGVIHRD
                     IKSDSILLTHDGRVKLSDFGFCAQVSKEVPRRKSLVGTPYWMAPELISRLPYGPEVDIW
                     SLGIMVIEMVDGEPPYFNEPPLKAMKMIRDNLPPRLKNLHKVSPSLKGFLDRLLVRDPA
                     QRATAAELLKHPFLAKAGPPASIVPLMRQNRTR"
     primer_bind     complement(1183..1201)
                     /label=PAK4-R
     primer_bind     complement(2011..2057)
                     /label=XmaI-PAK4-R
     polyA_signal    2084..2205
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2187..2205)
                     /label=SV40-POLYa
     promoter        2399..2756
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2399..2590
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     rep_origin      2607..2742
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2787..3824
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             3803..3811
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    3848..3896
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(4223..4811)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5011..5871)
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     polyA_signal    5976..6024
                     /note="synthetic polyadenylation signal"
     misc_feature    6038..6129
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
ORIGIN
        1 ggtacctgag ctcgctagct acattggaaa attttataca cgttctagct acattggaaa
       61 attttataca cgttctagct acattggaaa attttataca cgttctagct acattggaaa
      121 attttataca cgttctagct acattggaaa attttataca cgttagactc tagagggtat
      181 ataatggaag ctcgacttcc agcttggcaa tccggtactg ttggtaaaga attcgccacc
      241 atggtgagca agggcgagga gctgttcacc ggcgtggtgc ctatcctggt ggagctggac
      301 ggcgacgtga acggccacaa gttcagcgtg aggggcgagg gcgagggcga cgccaccaac
      361 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcctgtgcc ttggcctacc
      421 ctggtgacca ccctgggcta cggcgtggcc tgcttcgcca ggtaccctga ccacatgaag
      481 cagcacgact tcttcaagag cgccatgcct gagggctacg tgcaggagag gaccatcagc
      541 ttcaaggacg acggcaccta caagaccagg gccgaggtga agttcgaggg cgacaccctg
      601 gtgaacagga tcgtgctgaa gggcatcgac ttcaaggagg acggcaacat cctgggccac
      661 aagctggagt acaacttcaa cagccacaag gtgtacatca ccgccgacaa gcagaagaac
      721 ggcatcaagg ccaacttcaa gaccaggcac aacgtggagg acggcggcgt gcagctggcc
      781 gaccactacc agcagaacac ccctatcggc gacggccctg tgctgctgcc tgacaaccac
      841 tacctgagcc accagagcaa gctgagcaag gaccctaacg agaagaggga ccacatggtg
      901 ctgaaggaga gggtgaccgc cgccggcatc acccacgaca tggacgagct gtacaagtca
      961 ggtggatccg aagcagagga ctggacagca gccctactga acaggggtcg cagtcgccag
     1021 cccctggtac taggggacaa ttgctttgct gacttggtgc acaactggat ggagctgcct
     1081 gaggaattcc ccgccgcccc tgctgttcct gggccccctg gcccccgctc accacagcgg
     1141 gagccacagc gagtatccca tgagcagttc cgggctgccc tgcagctggt ggtggaccca
     1201 ggcgaccccc gctcctacct ggacaacttc atcaagattg gcgagggctc cacgggcatc
     1261 gtgtgcatcg ccaccgtgcg cagctcgggc aagctggtgg ccgtcaagaa gatggacctg
     1321 cgcaagcagc agaggcgcga gctgctcttc aacgaggtgg taatcatgag ggactaccag
     1381 cacgagaatg tggtggagat gtacaacagc tacctggtgg gggacgagct ctgggtggtc
     1441 atggagttcc tggaaggagg cgccctcacc gacatcgtca cccacaccag gatgaacgag
     1501 gagcagatcg cagccgtgtg ccttgcagtg ctgcaggccc tgtcggtgct ccacgcccag
     1561 ggcgtcatcc accgggacat caagagcgac tcgatcctgc tgacccatga tggcagggtg
     1621 aagctgtcag actttgggtt ctgcgcccag gtgagcaagg aagtgccccg aaggaagtcg
     1681 ctggtcggca cgccctactg gatggcccca gagctcatct cccgccttcc ctacgggcca
     1741 gaggtagaca tctggtcgct ggggataatg gtgattgaga tggtggacgg agagcccccc
     1801 tacttcaacg agccacccct caaagccatg aagatgattc gggacaacct gccaccccga
     1861 ctgaagaacc tgcacaaggt gtcgccatcc ctgaagggct tcctggaccg cctgctggtg
     1921 cgagaccctg cccagcgggc cacggcagcc gagctgctga agcacccatt cctggccaag
     1981 gcagggccgc ctgccagcat cgtgcccctc atgcgccaga accgcaccag atgacccggg
     2041 ttactctaga gtcggggcgg ccggccgctt cgagcagaca tgataagata cattgatgag
     2101 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     2161 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     2221 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
     2281 ctctacaaat gtggtaaaat cgataaggat ccgtttgcgt attgggcgct cttccgctga
     2341 tctgcgcagc accatggcct gaaataacct ctgaaagagg aacttggtta gctaccttct
     2401 gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     2461 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     2521 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     2581 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     2641 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     2701 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     2761 tcgattcttc tgacactagc gccaccatga agaagcccga actcaccgct accagcgttg
     2821 aaaaatttct catcgagaag ttcgacagtg tgagcgacct gatgcagttg tcggagggcg
     2881 aagagagccg agccttcagc ttcgatgtcg gcggacgcgg ctatgtactg cgggtgaata
     2941 gctgcgctga tggcttctac aaagaccgct acgtgtaccg ccacttcgcc agcgctgcac
     3001 tacccatccc cgaagtgttg gacatcggcg agttcagcga gagcctgaca tactgcatca
     3061 gtagacgcgc ccaaggcgtt actctccaag acctccccga aacagagctg cctgctgtgt
     3121 tacagcctgt cgccgaagct atggatgcta ttgccgccgc cgacctcagt caaaccagcg
     3181 gcttcggccc attcgggccc caaggcatcg gccagtacac aacctggcgg gatttcattt
     3241 gcgccattgc tgatccccat gtctaccact ggcagaccgt gatggacgac accgtgtccg
     3301 ccagcgtagc tcaagccctg gacgaactga tgctgtgggc cgaagactgt cccgaggtgc
     3361 gccacctcgt ccatgccgac ttcggcagca acaacgtcct gaccgacaac ggccgcatca
     3421 ccgccgtaat cgactggtcc gaagctatgt tcggggacag tcagtacgag gtggccaaca
     3481 tcttcttctg gcggccctgg ctggcttgca tggagcagca gactcgctac ttcgagcgcc
     3541 ggcatcccga gctggccggc agccctcgtc tgcgagccta catgctgcgc atcggcctgg
     3601 atcagctcta ccagagcctc gtggacggca acttcgacga tgctgcctgg gctcaaggcc
     3661 gctgcgatgc catcgtccgc agcggggccg gcaccgtcgg tcgcacacaa atcgctcgcc
     3721 ggagcgcagc cgtatggacc gacggctgcg tcgaggtgct ggccgacagc ggcaaccgcc
     3781 ggcccagtac acgaccgcgc gctaaggagg taggtcgagt ttaaactcta gaaccggtca
     3841 tggccgcaat aaaatatctt tattttcatt acatctgtgt gttggttttt tgtgtgttcg
     3901 aactagatgc tgtcgaccga tgcccttgag agccttcaac ccagtcagct ccttccggtg
     3961 ggcgcggggc atgactatcg tcgccgcact tatgactgtc ttctttatca tgcaactcgt
     4021 aggacaggtg ccggcagcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     4081 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     4141 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     4201 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4261 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4321 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4381 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4441 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4501 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4561 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     4621 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     4681 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     4741 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     4801 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     4861 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     4921 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     4981 gcggccgcaa atgctaaacc actgcagtgg ttaccagtgc ttgatcagtg aggcaccgat
     5041 ctcagcgatc tgcctatttc gttcgtccat agtggcctga ctccccgtcg tgtagatcac
     5101 tacgattcgt gagggcttac catcaggccc cagcgcagca atgatgccgc gagagccgcg
     5161 ttcaccggcc cccgatttgt cagcaatgaa ccagccagca gggagggccg agcgaagaag
     5221 tggtcctgct actttgtccg cctccatcca gtctatgagc tgctgtcgtg atgctagagt
     5281 aagaagttcg ccagtgagta gtttccgaag agttgtggcc attgctactg gcatcgtggt
     5341 atcacgctcg tcgttcggta tggcttcgtt caactctggt tcccagcggt caagccgggt
     5401 cacatgatca cccatattat gaagaaatgc agtcagctcc ttagggcctc cgatcgttgt
     5461 cagaagtaag ttggccgcgg tgttgtcgct catggtaatg gcagcactac acaattctct
     5521 taccgtcatg ccatccgtaa gatgcttttc cgtgaccggc gagtactcaa ccaagtcgtt
     5581 ttgtgagtag tgtatacggc gaccaagctg ctcttgcccg gcgtctatac gggacaacac
     5641 cgcgccacat agcagtactt tgaaagtgct catcatcggg aatcgttctt cggggcggaa
     5701 agactcaagg atcttgccgc tattgagatc cagttcgata tagcccactc ttgcacccag
     5761 ttgatcttca gcatctttta ctttcaccag cgtttcgggg tgtgcaaaaa caggcaagca
     5821 aaatgccgca aagaagggaa tgagtgcgac acgaaaatgt tggatgctca tactcgtcct
     5881 ttttcaatat tattgaagca tttatcaggg ttactagtac gtctctcaag gataagtaag
     5941 taatattaag gtacgggagg tattggacag gccgcaataa aatatcttta ttttcattac
     6001 atctgtgtgt tggttttttg tgtgaatcga tagtactaac atacgctctc catcaaaaca
     6061 aaacgaaaca aaacaaacta gcaaaatagg ctgtccccag tgcaagtgca ggtgccagaa
     6121 catttctctg gcctaactgg cc
//