LOCUS       sequence-318635-        7156 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  STELLA plasmid for amber suppression of N-terminally labelled 
            CoV2_ORF10.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7156)
  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 7156)
  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-*CoV2_ORF10
FEATURES             Location/Qualifiers
     source          1..7156
                     /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"
     primer_bind     complement(3624..3644)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    3686..3807
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3723..3742)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3777..3796
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   4570..4604
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(5095..5111)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5095..5111)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5108..5130)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5119..5135
                     /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(5143..5149,5150..5167,5168..5173))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5188..5209
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5326..5343)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5497..6085)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5577..5596)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6256..7047,7048..7116))
                     /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     6879..6898
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(7117..7156,1..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 ggggttaggg gtacataaat gtttttgcat tccccttcac
     3481 gatctatagt ttgttgctgt gtcggatgaa ctccagaaat tacattgctc aagttgatgt
     3541 ggtcaatttt aatctcactt gaccggcgac aatcaacctc tggattacaa aatttgtgaa
     3601 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     3661 atgcctttgt atcatgcgtt aactaaactt gtttattgca gcttataatg gttacaaata
     3721 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     3781 tttgtccaaa ctcatcaatg tatcttatca tgtctggaat tgactcaaat gatgtcaatt
     3841 agtctatcag aagctcatct ggtctccctt ccgggggaca agacatccct gtttaatatt
     3901 taaacagcag tgttcccaaa ctgggttctt atatcccttg ctctggtcaa ccaggttgca
     3961 gggtttcctg tcctcacagg aacgaagtcc ctaaagaaac agtggcagcc aggtttagcc
     4021 ccggaattga ctggattcct tttttagggc ccattggtat ggctttttcc ccgtatcccc
     4081 ccaggtgtct gcaggctcaa agagcagcga gaagcgttca gaggaaagcg atcccgtgcc
     4141 accttccccg tgcccgggct gtccccgcac gctgccggct cggggatgcg gggggagcgc
     4201 cggaccggag cggagccccg ggcggctcgc tgctgccccc tagcggggga gggacgtaat
     4261 tacatccctg ggggctttgg gggggggctg tccctgatat ctataacaag aaaatatata
     4321 tataataagt tatcacgtaa gtagaacatg aaataacaat ataattatcg tatgagttaa
     4381 atcttaaaag tcacgtaaaa gataatcatg cgtcattttg actcacgcgg tcgttatagt
     4441 tcaaaatcag tgacacttac cgcattgaca agcacgcctc acgggagctc caagcggcga
     4501 ctgagatgtc ctaaatgcac agcgacggat tcgcgctatt tagaaagaga gagcaatatt
     4561 tcaagaatgc atgcgtcaat tttacgcaga ctatctttct agggttaatc tagctgcatc
     4621 aggatcatat cgtcgggtct tttttccggc tcagtcatcg cccaagctgg cgctatctgg
     4681 gcatcgggga ggaagaagcc cgtgcctttt cccgcgaggt tgaagcggca tggaaagagt
     4741 ttgccgagga tgactgctgc tgcattgacg ttgagcgaaa acgcacgttt accatgatga
     4801 ttcgggaagg tgtggccatg cacgccttta acggtgaact gttcgttcag gccacctggg
     4861 ataccagttc gtcgcggctt ttccggacac agttccggat ggtcagcccg aagcgcatca
     4921 gcaacccgaa caataccggc gacagccgga actgccgtgc cggtgtgcag attaatgaca
     4981 gcggtgcggc gctgggatat tacgtcagcg aggacgggta tcctggctgg atgccgcaga
     5041 aatggacatg gataccccgt gagttacccg gcgggcgcgc ttggcgtaat catggtcata
     5101 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     5161 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     5221 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     5281 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     5341 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     5401 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     5461 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     5521 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     5581 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     5641 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     5701 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     5761 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     5821 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     5881 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     5941 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     6001 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     6061 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     6121 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     6181 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     6241 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     6301 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     6361 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     6421 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     6481 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     6541 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     6601 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     6661 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     6721 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     6781 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     6841 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     6901 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6961 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     7021 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     7081 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     7141 aagcatttat cagggt
//
