LOCUS       sequence-318675-        7780 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  STELLA plasmid for amber suppression of N-terminally labelled GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7780)
  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 7780)
  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_4x7SKPylT_EF1_Ub-*GFP
FEATURES             Location/Qualifiers
     source          1..7780
                     /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             3218..3445
                     /codon_start=1
                     /product="human ubiquitin"
                     /label=ubiquitin
                     /translation="MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIF
                     AGKQLEDGRTLSDYNIQKESTLHLVLRLRGG"
     CDS             4166..4183
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4248..4268)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    4310..4431
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4347..4366)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4401..4420
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   5194..5228
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(5719..5735)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5719..5735)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5732..5754)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5743..5759
                     /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(5767..5773,5774..5791,5792..5797))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5812..5833
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5950..5967)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6121..6709)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6201..6220)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6880..7671,7672..7740))
                     /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     7503..7522
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(7741..7780,1..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 gcgaattatg cagatcttcg tgaagactct
     3241 gactggtaag accatcaccc tcgaggttga gcccagtgac accatcgaga atgtcaaggc
     3301 aaagatccaa gataaggaag gcatccctcc tgaccagcag aggctgatct ttgctggaaa
     3361 acagctggaa gatgggcgca ccctgtctga ctacaacatc cagaaagagt ccaccctgca
     3421 cctggtgctc cgcttaagag ggggttagtc caagggcgag gaactgttca ccggcgtggt
     3481 gcccatcctg gtggaactgg atggcgacgt gaacggccac aagttctctg tgcggggaga
     3541 gggcgaaggc gacgccacaa atggcaagct gaccctgaag ttcatctgca ccaccggcaa
     3601 gctgcccgtg ccttggccta ccctcgtgac cacactgacc tacggcgtgc agtgcttcag
     3661 cagatacccc gaccatatga agcggcacga cttcttcaag agcgccatgc ccgagggcta
     3721 cgtgcaggaa cggaccatca gcttcaagga cgacggcacc tacaagacca gagccgaagt
     3781 gaagttcgag ggcgacaccc tcgtgaaccg gatcgagctg aagggcatcg atttcaaaga
     3841 ggacggcaac atcctgggcc acaagctgga gtacaacttc aacagccacc tggtgtacat
     3901 caccgccgac aagcagaaga acggcatcaa ggccaacttc aagatccggc acaacgtgga
     3961 agatggcagc gtgcagctgg ccgaccacta ccagcagaac acccccatcg gagatggccc
     4021 cgtgctgctg cccgacaacc actacctgag cacccagagc gtgctgagca aggaccccaa
     4081 cgagaagcgg gaccacatgg tgctgctgga attcgtgacc gccgctggca tcacccacgg
     4141 catggacgag ctgtacaagg gcagccacca tcaccatcac cattaaccgg cgacaatcaa
     4201 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     4261 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg cgttaactaa acttgtttat
     4321 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     4381 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     4441 gaattgactc aaatgatgtc aattagtcta tcagaagctc atctggtctc ccttccgggg
     4501 gacaagacat ccctgtttaa tatttaaaca gcagtgttcc caaactgggt tcttatatcc
     4561 cttgctctgg tcaaccaggt tgcagggttt cctgtcctca caggaacgaa gtccctaaag
     4621 aaacagtggc agccaggttt agccccggaa ttgactggat tcctttttta gggcccattg
     4681 gtatggcttt ttccccgtat ccccccaggt gtctgcaggc tcaaagagca gcgagaagcg
     4741 ttcagaggaa agcgatcccg tgccaccttc cccgtgcccg ggctgtcccc gcacgctgcc
     4801 ggctcgggga tgcgggggga gcgccggacc ggagcggagc cccgggcggc tcgctgctgc
     4861 cccctagcgg gggagggacg taattacatc cctgggggct ttgggggggg gctgtccctg
     4921 atatctataa caagaaaata tatatataat aagttatcac gtaagtagaa catgaaataa
     4981 caatataatt atcgtatgag ttaaatctta aaagtcacgt aaaagataat catgcgtcat
     5041 tttgactcac gcggtcgtta tagttcaaaa tcagtgacac ttaccgcatt gacaagcacg
     5101 cctcacggga gctccaagcg gcgactgaga tgtcctaaat gcacagcgac ggattcgcgc
     5161 tatttagaaa gagagagcaa tatttcaaga atgcatgcgt caattttacg cagactatct
     5221 ttctagggtt aatctagctg catcaggatc atatcgtcgg gtcttttttc cggctcagtc
     5281 atcgcccaag ctggcgctat ctgggcatcg gggaggaaga agcccgtgcc ttttcccgcg
     5341 aggttgaagc ggcatggaaa gagtttgccg aggatgactg ctgctgcatt gacgttgagc
     5401 gaaaacgcac gtttaccatg atgattcggg aaggtgtggc catgcacgcc tttaacggtg
     5461 aactgttcgt tcaggccacc tgggatacca gttcgtcgcg gcttttccgg acacagttcc
     5521 ggatggtcag cccgaagcgc atcagcaacc cgaacaatac cggcgacagc cggaactgcc
     5581 gtgccggtgt gcagattaat gacagcggtg cggcgctggg atattacgtc agcgaggacg
     5641 ggtatcctgg ctggatgccg cagaaatgga catggatacc ccgtgagtta cccggcgggc
     5701 gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     5761 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     5821 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     5881 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     5941 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     6001 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     6061 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     6121 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     6181 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     6241 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     6301 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     6361 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     6421 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     6481 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     6541 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     6601 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     6661 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     6721 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     6781 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     6841 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     6901 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     6961 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     7021 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     7081 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     7141 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     7201 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     7261 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     7321 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     7381 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     7441 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     7501 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     7561 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     7621 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     7681 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     7741 actcttcctt tttcaatatt attgaagcat ttatcagggt
//
