LOCUS       sequence-318685-        8384 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  STELLA plasmid for amber suppression of C-terminally labelled GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8384)
  AUTHORS   Lafranchi L, Schlesinger D, Kimler KJ, Elsasser SJ
  TITLE     Universal Single-Residue Terminal Labels for Fluorescent Live Cell 
            Imaging of Microproteins.
  JOURNAL   J Am Chem Soc. 2020 Nov 25;142(47):20080-20087. doi: 
            10.1021/jacs.0c09574. Epub 2020 Nov 11.
  PUBMED    33175524
REFERENCE   2  (bases 1 to 8384)
  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"; doi: 
            "10.1021/jacs.0c09574"; journalName: "J Am Chem Soc"; date: 
            "2020-11-25- 25"; volume: "142"; issue: "47"; pages: "20080-20087"
COMMENT     Sequence Label: pAS1_4xh7SKPylT_EF1_GFP*_Intein_DD
FEATURES             Location/Qualifiers
     source          1..8384
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(91..546)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(228..249)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     440..459
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     673..695
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     687..704
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     688..704
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   834..896
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        2687..2898
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             2911..3179
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             3945..3962
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             4470..4793
                     /codon_start=1
                     /product="destabilization domain that can be stabilized by 
                     Shield1 in the ProteoTuner(TM) system"
                     /label=DD-C
                     /note="FKBP12 mutant optimized for use as a C-terminal tag"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLGDGKKVDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQGAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     ELE"
     primer_bind     complement(4852..4872)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    4914..5035
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4951..4970)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5005..5024
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   5798..5832
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(6323..6339)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6323..6339)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6336..6358)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6347..6363
                     /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(6371..6377,6378..6395,6396..6401))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6416..6437
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6554..6571)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6725..7313)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6805..6824)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7484..8275,8276..8344))
                     /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     8107..8126
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(8345..8384,1..65))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
       61 ccgcgcacat ttccccgaaa agtgccacct aaattgtaag cgttaatatt ttgttaaaat
      121 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
      181 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
      241 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
      301 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
      361 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
      421 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
      481 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
      541 gcgcgtccca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
      601 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
      661 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgagcgc gcctcgttca
      721 ttcacgtttt tgaacccgtg gaggacgggc agactcgcgg tgcaaatgtg ttttacagcg
      781 tgatggagca gatgaagatg ctcgacacgc tgcagaacac gcagctagat taaccctaga
      841 aagataatca tattgtgacg tacgttaaag ataatcatgt gtaaaattga cgcatgtgtt
      901 ttatcggtct gtatatcgag gtttatttat taatttgaat agatattaag ttttattata
      961 tttacactta catactaata ataaattcaa caaacaattt atttatgttt atttatttat
     1021 taaaaaaaac aaaaactcaa aatttcttct ataaagtaac aaaactttta tgagggacag
     1081 ccccccccca aagcccccag ggatgtaatt acgtccctcc cccgctaggg ggcagcagcg
     1141 agccgcccgg ggctccgctc cggtccggcg ctccccccgc atccccgagc cggcagcgtg
     1201 cggggacagc ccgggcacgg ggaaggtggc acgggatcgc tttcctctga acgcttctcg
     1261 ctgctctttg agcctgcaga cacctggggg gatacgggga aaaggcctcc acggccaact
     1321 agtgaaaaac cgcacttgtc cggaaacccc gggaatctaa cccggctgaa cggatttaga
     1381 gtccgttcga tctacatgat caggtttccg aggtacccag gcggcgcaca agctatataa
     1441 acctgaagga aatctcaact ttacacttag gtcaagttac ttatcgtact agagcttcag
     1501 caggaaattt aactaaaatc taatttaacc agcatagcaa atatcattta ttcccaaaat
     1561 gctaaagttt gagataaacg gacttgattt ccggctgttt tgacactatc cagaatgcct
     1621 tgcagatggg tggggcatgc taaatactgc aggaaaaacc gcacttgtcc ggaaaccccg
     1681 ggaatctaac ccggctgaac ggatttagag tccgttcgat ctacatgatc aggtttccga
     1741 ggtacccagg cggcgcacaa gctatataaa cctgaaggaa atctcaactt tacacttagg
     1801 tcaagttact tatcgtacta gagcttcagc aggaaattta actaaaatct aatttaacca
     1861 gcatagcaaa tatcatttat tcccaaaatg ctaaagtttg agataaacgg acttgatttc
     1921 cggctgtttt gacactatcc agaatgcctt gcagatgggt ggggcatgct aaatactgca
     1981 ggaaaaaccg cacttgtccg gaaaccccgg gaatctaacc cggctgaacg gatttagagt
     2041 ccgttcgatc tacatgatca ggtttccgag gtacccaggc ggcgcacaag ctatataaac
     2101 ctgaaggaaa tctcaacttt acacttaggt caagttactt atcgtactag agcttcagca
     2161 ggaaatttaa ctaaaatcta atttaaccag catagcaaat atcatttatt cccaaaatgc
     2221 taaagtttga gataaacgga cttgatttcc ggctgttttg acactatcca gaatgccttg
     2281 cagatgggtg gggcatgcta aatactgcag gaaaaaccgc acttgtccgg aaaccccggg
     2341 aatctaaccc ggctgaacgg atttagagtc cgttcgatct acatgatcag gtttccgagg
     2401 tacccaggcg gcgcacaagc tatataaacc tgaaggaaat ctcaacttta cacttaggtc
     2461 aagttactta tcgtactaga gcttcagcag gaaatttaac taaaatctaa tttaaccagc
     2521 atagcaaata tcatttattc ccaaaatgct aaagtttgag ataaacggac ttgatttccg
     2581 gctgttttga cactatccag aatgccttgc agatgggtgg ggcatgctaa atactgcaga
     2641 actagtgctc tagaaaggat ctgcgatcgc tccggtgccc gtcagtgggc agagcgcaca
     2701 tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaacggg tgcctagaga
     2761 aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag
     2821 ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg
     2881 tttgccgcca gaacacagct gaagcttcga ggggctcgca tctctccttc acgcgcccgc
     2941 cgccctacct gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc tcccgcctgt
     3001 ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc gagaccgggc
     3061 ctttgtccgg cgctcccttg gagcctacct agactcagcc ggctctccac gctttgcctg
     3121 accctgcttg ctcaactcta cgtctttgtt tcgttttctg ttctgcgccg ttacagatcc
     3181 aagctgtgac cggcgcctac tctagagcta gcgaattcgc caccatgtcc aagggcgagg
     3241 aactgttcac cggcgtggtg cccatcctgg tggaactgga tggcgacgtg aacggccaca
     3301 agttctctgt gcggggagag ggcgaaggcg acgccacaaa tggcaagctg accctgaagt
     3361 tcatctgcac caccggcaag ctgcccgtgc cttggcctac cctcgtgacc acactgacct
     3421 acggcgtgca gtgcttcagc agataccccg accatatgaa gcggcacgac ttcttcaaga
     3481 gcgccatgcc cgagggctac gtgcaggaac ggaccatcag cttcaaggac gacggcacct
     3541 acaagaccag agccgaagtg aagttcgagg gcgacaccct cgtgaaccgg atcgagctga
     3601 agggcatcga tttcaaagag gacggcaaca tcctgggcca caagctggag tacaacttca
     3661 acagccacct ggtgtacatc accgccgaca agcagaagaa cggcatcaag gccaacttca
     3721 agatccggca caacgtggaa gatggcagcg tgcagctggc cgaccactac cagcagaaca
     3781 cccccatcgg agatggcccc gtgctgctgc ccgacaacca ctacctgagc acccagagcg
     3841 tgctgagcaa ggaccccaac gagaagcggg accacatggt gctgctggaa ttcgtgaccg
     3901 ccgctggcat cacccacggc atggacgagc tgtacaaggg cagccaccat caccatcacc
     3961 attagtgcct cagttttggc accgaaattt taaccgttga gtacggccca ttgcccattg
     4021 gcaaaattgt gagtgaagaa attaattgtt ctgtgtacag tgttgatcca gaagggagag
     4081 tttacaccca ggcgatcgcc caatggcatg accggggaga gcaggaagta ttggaatatg
     4141 aattggaaga tggttcagta atccgagcta cctctgacca ccgcttttta accaccgatt
     4201 atcaactgtt ggcgatcgaa gaaatttttg ctaggcaact ggacttgttg actttagaaa
     4261 atattaagca aactgaagaa gctcttgaca accatcgtct tccctttcca ttacttgacg
     4321 ctgggacaat taaaatggtt aaagttatcg gtcgtcgttc cctcggagtg caaagaatat
     4381 ttgatattgg tcttccccaa gaccataatt ttctgctagc caatggggcg atcgccgccg
     4441 cctgcttcaa cgaattcgaa ttcctcgagg gagtgcaggt ggaaaccatc tccccaggtg
     4501 acgggcgcac cttccccaag cgcggccaga cctgcgtggt gcactacacc gggatgcttg
     4561 gagatggaaa gaaagttgac tcctcccggg acagaaacaa gccctttaag tttatgctag
     4621 gcaagcagga ggtgatccga ggctgggaag aaggggttgc ccagatgagt gtgggtcagg
     4681 gagccaaact gactatatct ccagattatg cctatggtgc cactgggcac ccaggcatca
     4741 tcccaccaca tgccactctc gtcttcgatg tggagcttct agaactggaa tgatcgacaa
     4801 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     4861 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgcgttaa ctaaacttgt
     4921 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     4981 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     5041 tctggaattg actcaaatga tgtcaattag tctatcagaa gctcatctgg tctcccttcc
     5101 gggggacaag acatccctgt ttaatattta aacagcagtg ttcccaaact gggttcttat
     5161 atcccttgct ctggtcaacc aggttgcagg gtttcctgtc ctcacaggaa cgaagtccct
     5221 aaagaaacag tggcagccag gtttagcccc ggaattgact ggattccttt tttagggccc
     5281 attggtatgg ctttttcccc gtatcccccc aggtgtctgc aggctcaaag agcagcgaga
     5341 agcgttcaga ggaaagcgat cccgtgccac cttccccgtg cccgggctgt ccccgcacgc
     5401 tgccggctcg gggatgcggg gggagcgccg gaccggagcg gagccccggg cggctcgctg
     5461 ctgcccccta gcgggggagg gacgtaatta catccctggg ggctttgggg gggggctgtc
     5521 cctgatatct ataacaagaa aatatatata taataagtta tcacgtaagt agaacatgaa
     5581 ataacaatat aattatcgta tgagttaaat cttaaaagtc acgtaaaaga taatcatgcg
     5641 tcattttgac tcacgcggtc gttatagttc aaaatcagtg acacttaccg cattgacaag
     5701 cacgcctcac gggagctcca agcggcgact gagatgtcct aaatgcacag cgacggattc
     5761 gcgctattta gaaagagaga gcaatatttc aagaatgcat gcgtcaattt tacgcagact
     5821 atctttctag ggttaatcta gctgcatcag gatcatatcg tcgggtcttt tttccggctc
     5881 agtcatcgcc caagctggcg ctatctgggc atcggggagg aagaagcccg tgccttttcc
     5941 cgcgaggttg aagcggcatg gaaagagttt gccgaggatg actgctgctg cattgacgtt
     6001 gagcgaaaac gcacgtttac catgatgatt cgggaaggtg tggccatgca cgcctttaac
     6061 ggtgaactgt tcgttcaggc cacctgggat accagttcgt cgcggctttt ccggacacag
     6121 ttccggatgg tcagcccgaa gcgcatcagc aacccgaaca ataccggcga cagccggaac
     6181 tgccgtgccg gtgtgcagat taatgacagc ggtgcggcgc tgggatatta cgtcagcgag
     6241 gacgggtatc ctggctggat gccgcagaaa tggacatgga taccccgtga gttacccggc
     6301 gggcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     6361 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     6421 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     6481 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     6541 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     6601 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     6661 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     6721 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     6781 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     6841 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6901 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     6961 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     7021 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     7081 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     7141 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     7201 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     7261 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     7321 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     7381 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     7441 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     7501 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     7561 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     7621 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     7681 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     7741 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     7801 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     7861 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     7921 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     7981 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     8041 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     8101 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     8161 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     8221 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     8281 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     8341 tcatactctt cctttttcaa tattattgaa gcatttatca gggt
//
