LOCUS       sequence-130228-        6027 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Aurein1.2_+36GFP-Cys expression in E.Coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6027)
  AUTHORS   Li M, Tao Y, Shu Y, LaRochelle JR, Steinauer A, Thompson D, 
            Schepartz A, Chen ZY, Liu DR
  TITLE     Discovery and Characterization of a Peptide That Enhances Endosomal 
            Escape of Delivered Proteins in Vitro and in Vivo.
  JOURNAL   J Am Chem Soc. 2015 Nov 11;137(44):14084-93. doi: 
            10.1021/jacs.5b05694. Epub 2015 Oct 30.
  PUBMED    26465072
REFERENCE   2  (bases 1 to 6027)
  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.5b05694"; journalName: "J Am Chem Soc"; date: 
            "2015-11-11- 11"; volume: "137"; issue: "44"; pages: "14084-93"
COMMENT     Sequence Label: Aurein1.2_(pos36GFP)-Cys
FEATURES             Location/Qualifiers
     source          1..6027
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    2..26
                     /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)."
     RBS             41..63
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             937..954
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             973..1017
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1055..1073)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1069..1116
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1153..1608
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1240..1259)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1450..1471
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(1723..2514,2515..2583))
                     /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     2346..2365
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2584..2676)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2757..3345
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3246..3265
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3499..3516
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3531..3673
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3599..3618)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3759..3781
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3775..3966)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4538..5620)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     5582..5601
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5621..5698)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5904..5923
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     join(6011..6027,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(6011..6027,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gggaattgtg agcggataac aattcccctc tagaaataat tttgtttaac tttaagaagg
       61 agatatcata tgggcctgtt tgatattatt aaaaaaattg cggaaagctt tgctagcggt
      121 ggtagcggtg gttctggtgg ttctggtggt agcggcggta gcggtggtag cggtggtagc
      181 ggtggcagcg gcggttctag caaaggtgaa cgtctgtttc gtggtaaagt accgatctta
      241 gtggaattaa agggcgacgt gaacggtcat aaatttagcg tgcgcggcaa aggcaaaggt
      301 gacgctaccc gtggtaaatt gaccctgaag tttatttgca caacaggcaa attacccgtt
      361 ccgtggccca ccttagtgac caccctgacc tatggcgttc agtgcttcag tcgttaccct
      421 aaacacatga aacgtcacga ttttttcaaa tcagccatgc ctaaaggata tgttcaagag
      481 cgtacaatca gcttcaagaa ggatggcaaa tataaaacgc gtgcggaagt gaaatttgaa
      541 ggccgcacat tagtaaatcg tatcaaactg aaaggtcgtg acttcaaaga aaaaggcaac
      601 attttaggcc ataaactgcg ttataacttt aattctcata aggtgtatat tacggccgat
      661 aaacgcaaga atggtatcaa ggcaaaattc aaaattcgcc ataacgtgaa agacggcagc
      721 gttcaattag cggatcatta tcaacaaaac acgccgattg gtcgcgggcc tgtactgtta
      781 cctcgcaacc actacctgag cacccgttct aaacttagca aagatccgaa agaaaaacgc
      841 gatcacatgg ttctgttaga attcgtgacc gctgcaggca ttaagcacgg acgcgacgaa
      901 cgctacaaga ccggtggtag cggatgcgga ggatcccatc accaccacca tcactaatga
      961 ctcgagtctg gtaaagaaac cgctgctgcg aaatttgaac gccagcacat ggactcgtct
     1021 actagcgcag cttaattaac ctaggctgct gccaccgctg agcaataact agcataaccc
     1081 cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga
     1141 ttggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     1201 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     1261 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     1321 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     1381 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     1441 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     1501 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     1561 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttct ggcggcacga
     1621 tggcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     1681 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     1741 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     1801 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     1861 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     1921 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     1981 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     2041 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     2101 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     2161 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     2221 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     2281 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     2341 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     2401 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     2461 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     2521 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     2581 catactcttc ctttttcaat catgattgaa gcatttatca gggttattgt ctcatgagcg
     2641 gatacatatt tgaatgtatt tagaaaaata aacaaatagg tcatgaccaa aatcccttaa
     2701 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     2761 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     2821 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     2881 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     2941 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     3001 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     3061 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     3121 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     3181 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     3241 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     3301 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     3361 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     3421 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     3481 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg
     3541 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatatggtgc actctcagta
     3601 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg
     3661 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     3721 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     3781 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc
     3841 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag
     3901 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt
     3961 ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa
     4021 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg
     4081 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg
     4141 tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc
     4201 tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta
     4261 cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca
     4321 gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc
     4381 ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgctagtca tgccccgcgc
     4441 ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc
     4501 ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     4561 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     4621 tattgggcgc cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct
     4681 tcaccgcctg gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc
     4741 gaaaatcctg tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt
     4801 cgtatcccac taccgagatg tccgcaccaa cgcgcagccc ggactcggta atggcgcgca
     4861 ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat
     4921 tcagcatttg catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg
     4981 ctatcggctg aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg
     5041 ccgagacaga acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca
     5101 gatgctccac gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg
     5161 tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa
     5221 tggcatcctg gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa
     5281 gattgtgcac cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca
     5341 cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt
     5401 gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt
     5461 gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg
     5521 ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac
     5581 cggcatactc tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac
     5641 tctcttccgg gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggtgtccg
     5701 ggatctcgac gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg
     5761 aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc gcccaacagt
     5821 cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag
     5881 tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct
     5941 gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat cgatctcgat
     6001 cccgcgaaat taatacgact cactata
//
