LOCUS       IronFIST        4812 bp DNA     circular SYN 21-JUL-2025
DEFINITION  .
ACCESSION   VB240509-1013rcf
VERSION     .
KEYWORDS    IronFist
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4812)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 21, 2025 from SnapGene Viewer 6.2.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4812
                     /note="color: #ffffff"
                     /organism="synthetic DNA construct"
     misc_feature    22..27
                     /label=BamHI
                     /note="/__SeqFeature__=True"
                     /note="/__level__=0"
                     /note="/name=BamHI"
                     /note="/official_designation=BamHI"
                     /note="/uuid=2e24d26a052f4a8f9539513b7d1f974d"
                     /note="color: #d05c0a; direction: RIGHT"
     misc_feature    28..532
                     /label=hPGK
                     /note="/__SeqFeature__=True"
                     /note="/__level__=0"
                     /note="/application_notes=Medium-strength promoter."
                     /note="/name=hPGK"
                     /note="/official_designation=hPGK promoter"
                     /note="/uuid=a105ee8f48814e769c9a41c38cfff41e"
                     /note="color: #fd3434; direction: RIGHT"
     misc_feature    533..538
                     /label=Kozak
                     /label=nonstandard type:  Miscellaneous
                     /note="/Application Notes=Facilitates translation 
                     initiation ofATG start codon downstream of the Kozak 
                     sequence."
                     /note="/Full Name=Kozak"
                     /note="/References=Nucleic Acids Res. 9:5233 (1981)"
                     /note="/__SeqFeature__=True"
                     /note="/__level__=0"
                     /note="/application_notes=Facilitates translation 
                     initiation ofATG start codon downstream of the Kozak 
                     sequence."
                     /note="/description=Kozak translation initiation sequence"
                     /note="/full_name=Kozak"
                     /note="/name=Kozak"
                     /note="/official_designation=Kozak"
                     /note="/uuid=5f18b25a1234443494a618363f09f052"
                     /note="color: #46c6ef; direction: RIGHT"
     CDS             539..1738
                     /label={Hr-mNeonGreen kpn1}
                     /note="/__SeqFeature__=True"
                     /note="/__level__=0"
                     /note="/name={Hr-mNeonGreen kpn1}"
                     /note="/official_designation={Hr-mNeonGreen kpn1}"
                     /note="/uuid=0fb06c1447db44449b88d56e3bbe548c"
                     /note="color: #eda481; direction: RIGHT"
     misc_feature    1739..1801
                     /label=T2A
                     /label=nonstandard type:  Linker
                     /note="/Application Notes=Causes co-translational cleavage 
                     of theencoded polypeptide. Multiple proteins can be made 
                     from a polycistronic transcript containing multiple ORFs 
                     separated by 2A. P2A and T2A have higher cleavage 
                     efficiency compared to other 2As."
                     /note="/Full Name=Thosea asigna virus 2A peptide"
                     /note="/References=PLoS One. 6:e18556(2011)"
                     /note="/Subtype=Read-through linker"
                     /note="/__SeqFeature__=True"
                     /note="/__level__=0"
                     /note="/application_notes=Causes co-translational cleavage 
                     of theencoded polypeptide. Multiple proteins can be made 
                     from a polycistronic transcript containing multiple ORFs 
                     separated by 2A. P2A and T2A have higher cleavage 
                     efficiency compared to other 2As."
                     /note="/description=Self-cleaving 2A peptide from Thosea 
                     asignavirus"
                     /note="/full_name=Thosea asigna virus 2A peptide"
                     /note="/name=T2A"
                     /note="/official_designation=T2A"
                     /note="/uuid=995274d0c39f4cbdbab66fb78aa938d6"
                     /note="color: #da3bb5; direction: RIGHT"
     CDS             1802..2515
                     /label={mCherry}
                     /note="/__SeqFeature__=True"
                     /note="/__level__=0"
                     /note="/name={mCherry}"
                     /note="/official_designation={mCherry}"
                     /note="/uuid=de2b71690ccd4ad0b3894d6e8a9449b5"
                     /note="color: #e4b930; direction: RIGHT"
     polyA_signal    2560..2781
                     /label=SV40 late pA
                     /note="/Application Notes=Allows transcription termination 
                     andpolyadenylation of mRNA transcribed by Pol II RNA 
                     polymerase."
                     /note="/References=Mol. Cell. Biol.9:4248 (1989)"
                     /note="/__SeqFeature__=True"
                     /note="/__level__=0"
                     /note="/application_notes=Allows transcription termination 
                     andpolyadenylation of mRNA transcribed by Pol II RNA 
                     polymerase."
                     /note="/description=Simian virus 40 late polyadenylation 
                     signal"
                     /note="/name=SV40 late pA"
                     /note="/official_designation=SV40 late pA"
                     /note="/uuid=d23bf533e0924dfcb932fc3ddd77e791"
                     /note="color: #7a00b5; direction: RIGHT"
     rep_origin      complement(2977..3565)
                     /direction=LEFT
                     /label=pUC ori
                     /note="/Application Notes=Facilitates plasmid replication 
                     in E.coli; regulates high-copy plasmid number (500-700)."
                     /note="/Full Name=pUC origin of replication"
                     /note="/References=Mol Microbiol. 6:3385 (1992)"
                     /note="/Subtype=E. coli replication_origin"
                     /note="/__SeqFeature__=True"
                     /note="/__level__=0"
                     /note="/application_notes=Facilitates plasmid replication 
                     in E.coli; regulates high-copy plasmid number (500-700)."
                     /note="/description=pUC origin of replication"
                     /note="/full_name=pUC origin of replication"
                     /note="/name=pUC ori"
                     /note="/official_designation=pUC ori"
                     /note="/uuid=41c627938a364ebfa136b55444385c5b"
                     /note="color: #05696d"
     CDS             complement(3736..4596)
                     /label=Ampicillin
                     /note="/Application Notes=Allows E. coli to be resistant 
                     toampiciIIin."
                     /note="/Full Name=Ampicillin resistance gene"
                     /note="/References=Proc Natl Acad Sci U S A. 75: 3737 
                     (1978)"
                     /note="/Selection Drug=Ampicillin "
                     /note="/Subtype=E. coli selection marker"
                     /note="/Usage=Positive selection"
                     /note="/__SeqFeature__=True"
                     /note="/__level__=0"
                     /note="/application_notes=Allows E. coli to be resistant 
                     toampicillin."
                     /note="/description=AmpiciIIin resistance gene"
                     /note="/full_name=Ampicillin resistance gene"
                     /note="/name=Ampicillin"
                     /note="/official_designation=Ampicillin"
                     /note="/uuid=0bc7d58885794858b29e98fe171ff1d5"
                     /note="color: #f54c6b; direction: LEFT"
ORIGIN
        1 caactttgta tagaaaagtt gggatccggg ttgcgccttt tccaaggcag ccctgggttt
       61 gcgcagggac gcggctgctc tgggcgtggt tccgggaaac gcagcggcgc cgaccctggg
      121 tctcgcacat tcttcacgtc cgttcgcagc gtcacccgga tcttcgccgc tacccttgtg
      181 ggccccccgg cgacgcttcc tgctccgccc ctaagtcggg aaggttcctt gcggttcgcg
      241 gcgtgccgga cgtgacaaac ggaagccgca cgtctcacta gtaccctcgc agacggacag
      301 cgccagggag caatggcagc gcgccgaccg cgatgggctg tggccaatag cggctgctca
      361 gcagggcgcg ccgagagcag cggccgggaa ggggcggtgc gggaggcggg gtgtggggcg
      421 gtagtgtggg ccctgttcct gcccgcgcgg tgttccgcat tctgcaagcc tccggagcgc
      481 acgtcggcag tcggctccct cgttgaccga atcaccgacc tctctcccca gggccaccat
      541 ggcgcccttt cctgaagaag tggacgtctt caccgcccca cactggcgga tgaagcagct
      601 ggtggggctc tactgcgaca agctttctaa aaccaatttt tccaacaaca acgatttccg
      661 tgctcttctg cagtctttgt atgctacttt caaggagttc aaaatgcatg agcagattga
      721 aaatgaatac attattggtt tgcttcaaca acgcagccag accatttata atgtacattc
      781 tgacaataaa ctctccgaga tgcttagcct ctttgaaaag ggactgaaga atgttaagaa
      841 tgaatatgaa cagttaaatt atgcaaaaca actgaaagag agattggagg cttttacaag
      901 agattttctt cctcacatga aagaggaaga ggaggttttt cagcccatgt taatggaata
      961 ttttacctat gaagagctta aggatattaa aaagaaagtg attgcacaac actgctctgg
     1021 cggcggcgtg agtaaggggg aggaagataa catggcctca cttccagcga ctcacgaact
     1081 tcacattttt gggtccatca atggcgtgga tttcgatatg gtagggcaag ggactggcaa
     1141 ccctaacgac ggttacgaag aactcaatct gaaatcaacg aaaggggatt tgcaattttc
     1201 cccatggata ctcgtccccc atataggcta tggattccac caatatctgc cgtatccaga
     1261 cggaatgtct ccgtttcaag ccgcaatggt agacggtagc ggttaccagg tccaccggac
     1321 gatgcaattc gaagatgggg cctccctgac agtaaattat agatatacct atgaagggag
     1381 tcacataaag ggagaagctc aggtcaaagg aacgggtttc ccggctgacg gaccggtgat
     1441 gaccaattca ctgaccgccg ccgattggtg ccgctctaaa aaaacttacc caaatgataa
     1501 gactatcatt tcaactttta aatggagtta cactacgggg aatggtaaga ggtacaggtc
     1561 aacagccagg acaacttaca catttgccaa gccaatggcg gcgaactatt tgaaaaatca
     1621 gccaatgtac gttttccgaa aaaccgaatt gaaacattct aagaccgagc ttaatttcaa
     1681 ggaatggcaa aaagcattta cggatgtgat ggggatggat gagctttaca agggtaccgg
     1741 aagcggagag ggcaggggaa gtcttctaac atgcggggac gtggaggaaa atcccggccc
     1801 catggtctcc aaaggagagg aagacaatat ggccattatc aaagagttta tgcgatttaa
     1861 agtgcacatg gagggaagcg tcaacgggca tgagtttgag attgaggggg aaggagaggg
     1921 aagaccttat gaagggaccc agactgccaa actgaaagtc accaagggcg gacccctgcc
     1981 atttgcttgg gacattctga gcccacaatt catgtatggg tccaaagcct atgttaaaca
     2041 cccggcagac atacctgact acctcaagct ctcctttcct gaggggttta aatgggagag
     2101 ggtgatgaac tttgaagatg gaggcgtcgt gacagtaact caggatagca gcctgcaaga
     2161 cggggagttt atttacaagg ttaaattgcg cggaaccaac tttcccagcg atggacccgt
     2221 gatgcagaag aaaacaatgg gctgggaagc atcatccgaa agaatgtatc ccgaagatgg
     2281 cgcgttgaag ggggaaatca aacaaaggct caagttgaaa gatggagggc actatgacgc
     2341 tgaggtcaaa accacgtaca aggctaaaaa accggtacaa ctgcccggag catataatgt
     2401 taatattaag ctggacatca ccagccacaa tgaggattac acaatagtgg aacagtatga
     2461 aagagccgag ggaagacact caactggggg catggatgaa ctgtataaat gataaaccca
     2521 gctttcttgt acaaagtggt gatggccggc cgcttcgagc agacatgata agatacattg
     2581 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     2641 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     2701 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt
     2761 aaaacctcta caaatgtggt agcggccgcg gcgctcttcc gcttcctcgc tcactgactc
     2821 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     2881 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     2941 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3001 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3061 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     3121 taccggatac ctgtccgcct ttctctcttc gggaagcgtg gcgctttctc atagctcacg
     3181 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     3241 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     3301 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     3361 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     3421 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     3481 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3541 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3601 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     3661 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     3721 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     3781 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     3841 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     3901 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     3961 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     4021 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     4081 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     4141 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     4201 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     4261 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     4321 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     4381 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     4441 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     4501 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     4561 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     4621 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     4681 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     4741 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcggcg
     4801 cgccgcggcc gc
//
