LOCUS       sequence-318631-        7261 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  STELLA plasmid for amber suppression of N-terminally labelled 
            CoV2_ORF9c/14.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7261)
  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 7261)
  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_ORF9c/14
FEATURES             Location/Qualifiers
     source          1..7261
                     /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(3729..3749)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    3791..3912
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3828..3847)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3882..3901
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   4675..4709
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(5200..5216)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5200..5216)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5213..5235)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5224..5240
                     /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(5248..5254,5255..5272,5273..5278))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5293..5314
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5431..5448)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5602..6190)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5682..5701)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6361..7152,7153..7221))
                     /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     6984..7003
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(7222..7261,1..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 ggggttagct tcaatcctgc tacaacttcc tcaaggaaca
     3481 acactgtcaa aaggcatcta cccaaaaagg ggctgaggcc gcagttaaac ctctgttggt
     3541 accacatcac gtcgtagcta cggtacaaga aattcagctc caagcagcag ttggcgaact
     3601 gctcctgttg gaatggctcg ctatggctgt catgctcctt cttctctgtt gctgcttgac
     3661 agattgaccg gcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     3721 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     3781 gcgttaacta aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     3841 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     3901 caatgtatct tatcatgtct ggaattgact caaatgatgt caattagtct atcagaagct
     3961 catctggtct cccttccggg ggacaagaca tccctgttta atatttaaac agcagtgttc
     4021 ccaaactggg ttcttatatc ccttgctctg gtcaaccagg ttgcagggtt tcctgtcctc
     4081 acaggaacga agtccctaaa gaaacagtgg cagccaggtt tagccccgga attgactgga
     4141 ttcctttttt agggcccatt ggtatggctt tttccccgta tccccccagg tgtctgcagg
     4201 ctcaaagagc agcgagaagc gttcagagga aagcgatccc gtgccacctt ccccgtgccc
     4261 gggctgtccc cgcacgctgc cggctcgggg atgcgggggg agcgccggac cggagcggag
     4321 ccccgggcgg ctcgctgctg ccccctagcg ggggagggac gtaattacat ccctgggggc
     4381 tttggggggg ggctgtccct gatatctata acaagaaaat atatatataa taagttatca
     4441 cgtaagtaga acatgaaata acaatataat tatcgtatga gttaaatctt aaaagtcacg
     4501 taaaagataa tcatgcgtca ttttgactca cgcggtcgtt atagttcaaa atcagtgaca
     4561 cttaccgcat tgacaagcac gcctcacggg agctccaagc ggcgactgag atgtcctaaa
     4621 tgcacagcga cggattcgcg ctatttagaa agagagagca atatttcaag aatgcatgcg
     4681 tcaattttac gcagactatc tttctagggt taatctagct gcatcaggat catatcgtcg
     4741 ggtctttttt ccggctcagt catcgcccaa gctggcgcta tctgggcatc ggggaggaag
     4801 aagcccgtgc cttttcccgc gaggttgaag cggcatggaa agagtttgcc gaggatgact
     4861 gctgctgcat tgacgttgag cgaaaacgca cgtttaccat gatgattcgg gaaggtgtgg
     4921 ccatgcacgc ctttaacggt gaactgttcg ttcaggccac ctgggatacc agttcgtcgc
     4981 ggcttttccg gacacagttc cggatggtca gcccgaagcg catcagcaac ccgaacaata
     5041 ccggcgacag ccggaactgc cgtgccggtg tgcagattaa tgacagcggt gcggcgctgg
     5101 gatattacgt cagcgaggac gggtatcctg gctggatgcc gcagaaatgg acatggatac
     5161 cccgtgagtt acccggcggg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     5221 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     5281 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     5341 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     5401 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     5461 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     5521 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     5581 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     5641 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     5701 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     5761 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     5821 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     5881 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     5941 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     6001 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     6061 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     6121 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     6181 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     6241 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     6301 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     6361 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     6421 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     6481 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     6541 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     6601 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     6661 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     6721 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     6781 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     6841 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     6901 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     6961 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     7021 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     7081 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     7141 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     7201 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     7261 t
//
