LOCUS       sequence-492803-        5456 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  His-tagged superfolder GFP C-terminally fused to C-terminal LCGC14 
            split intein fragment.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5456)
  AUTHORS   Humberg C, Kroger J, Pietrokovski S, Mootz HD
  TITLE     Three-Segment Protein Labeling Using a Highly Efficient and 
            Cysteine-Less Split Intein Identified with Computational Prediction 
            of Aggregation Properties.
  JOURNAL   Angew Chem Int Ed Engl. 2025 Oct 23:e202515821. doi: 
            10.1002/anie.202515821.
  PUBMED    41131872
REFERENCE   2  (bases 1 to 5456)
  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"; journalName: "Angew Chem 
            Int Ed Engl. 2025 Oct 23:e202515821. doi: 10.1002/anie.202515821."
COMMENT     Sequence Label: pBAD_LcgcC-GFP-H6
FEATURES             Location/Qualifiers
     source          1..5456
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(38..55)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      208..294
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      386..413
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        432..523
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(524..592,593..1384)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(742..761)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1426..1881
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1513..1532)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1723..1744
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      1992..2580
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2481..2500
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2734..2751
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2766..2906
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2832..2851)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2992..3014
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3152..3171
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(3351..4229)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        4256..4540
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     4463..4482
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             4567..4589
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5437..5454
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 agcttggctg ttttggcgga tgagagaaga ttttcagcct gatacagatt aaatcagaac
       61 gcagaagcgg tctgataaaa cagaatttgc ctggcggcag tagcgcggtg gtcccacctg
      121 accccatgcc gaactcagaa gtgaaacgcc gtagcgccga tggtagtgtg gggtctcccc
      181 atgcgagagt agggaactgc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg
      241 gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg
      301 ggagcggatt tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg acgcccgcca
      361 taaactgcca ggcatcaaat taagcagaag gccatcctga cggatggcct ttttgcgttt
      421 ctacaaactc ttttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
      481 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
      541 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
      601 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
      661 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
      721 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca
      781 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
      841 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
      901 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
      961 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     1021 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac
     1081 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     1141 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     1201 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     1261 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     1321 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     1381 gtaactgtca gaccaagttt actcatatat actttagatt gatttacgcg ccctgtagcg
     1441 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     1501 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     1561 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     1621 tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga
     1681 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     1741 ctggaacaac actcaaccct atctcgggct attcttttga tttataaggg attttgccga
     1801 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     1861 aaatattaac gtttacaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     1921 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     1981 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     2041 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     2101 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
     2161 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     2221 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     2281 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     2341 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     2401 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     2461 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     2521 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     2581 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     2641 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     2701 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     2761 gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccggatatgg
     2821 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat
     2881 cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct
     2941 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     3001 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagaag gagatggcgc
     3061 ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga
     3121 gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa
     3181 ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atctgctcat
     3241 gtttgacagc ttatcatcga tgcataatgt gcctgtcaaa tggacgaagc agggattctg
     3301 caaaccctat gctactccgt caagccgtca attgtctgat tcgttaccaa ttatgacaac
     3361 ttgacggcta catcattcac tttttcttca caaccggcac ggaactcgct cgggctggcc
     3421 ccggtgcatt ttttaaatac ccgcgagaaa tagagttgat cgtcaaaacc aacattgcga
     3481 ccgacggtgg cgataggcat ccgggtggtg ctcaaaagca gcttcgcctg gctgatacgt
     3541 tggtcctcgc gccagcttaa gacgctaatc cctaactgct ggcggaaaag atgtgacaga
     3601 cgcgacggcg acaagcaaac atgctgtgcg acgctggcga tatcaaaatt gctgtctgcc
     3661 aggtgatcgc tgatgtactg acaagcctcg cgtacccgat tatccatcgg tggatggagc
     3721 gactcgttaa tcgcttccat gcgccgcagt aacaattgct caagcagatt tatcgccagc
     3781 agctccgaat agcgcccttc cccttgcccg gcgttaatga tttgcccaaa caggtcgctg
     3841 aaatgcggct ggtgcgcttc atccgggcga aagaaccccg tattggcaaa tattgacggc
     3901 cagttaagcc attcatgcca gtaggcgcgc ggacgaaagt aaacccactg gtgataccat
     3961 tcgcgagcct ccggatgacg accgtagtga tgaatctctc ctggcgggaa cagcaaaata
     4021 tcacccggtc ggcaaacaaa ttctcgtccc tgatttttca ccaccccctg accgcgaatg
     4081 gtgagattga gaatataacc tttcattccc agcggtcggt cgataaaaaa atcgagataa
     4141 ccgttggcct caatcggcgt taaacccgcc accagatggg cattaaacga gtatcccggc
     4201 agcaggggat cattttgcgc ttcagccata cttttcatac tcccgccatt cagagaagaa
     4261 accaattgtc catattgcat cagacattgc cgtcactgcg tcttttactg gctcttctcg
     4321 ctaaccaaac cggtaacccc gcttattaaa agcattctgt aacaaagcgg gaccaaagcc
     4381 atgacaaaaa cgcgtaacaa aagtgtctat aatcacggca gaaaagtcca cattgattat
     4441 ttgcacggcg tcacactttg ctatgccata gcatttttat ccataagatt agcggatcct
     4501 acctgacgct ttttatcgca actctctact gtttctccat acccgttttt ttgggctaga
     4561 aataattttg tttaacttta agaaggagat atacatatgg tggagaaatt gaagattcaa
     4621 aagattgaga aactggaaga ctttgacaac gagtatgtct atgacatctc cgtggataaa
     4681 gagacgccat atttcttcgg caataacatc cttgttcata acagcgctta tggtaccagc
     4741 aaaggcgaag aactgtttac cggcgtggtg ccgattctgg ttgaactgga tggcgatgtt
     4801 aatggccata agtttagcgt gcgcggcgaa ggcgaaggcg atgcgaccaa cggcaaactg
     4861 accctgaagt ttatttgcac caccggcaaa ctgccggtgc cgtggccgac cctggtgacc
     4921 accctgacct atggcgtgca gtgctttagc cgctatccgg atcacatgaa acgccatgat
     4981 ttctttaagt ccgcgatgcc ggaaggctat gttcaggaac gcaccattag ctttaaagat
     5041 gatggcacct ataagacccg cgcggaagtg aaatttgaag gcgataccct ggtgaaccgc
     5101 attgaactga aaggcattga cttcaaagaa gatggcaaca ttctgggcca taaactggag
     5161 tacaatttca acagccataa cgtgtatatt accgcggaca aacagaagaa cggcatcaaa
     5221 gcgaatttca aaatccgcca taatgtggaa gatggcagcg tgcagctggc ggatcactat
     5281 cagcagaata ccccgatcgg cgatggcccg gtgctgctgc cggacaatca ctacctgagc
     5341 acccagtccg tgctgagcaa agatccgaat gagaaacgcg atcacatggt tctgctggag
     5401 tttgtgaccg cggcgggtat cactcatggc ggcagccatc accatcacca tcacta
//
