LOCUS       sequence-138848-        7122 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Aurein1.2+36GFP-Cre fusion expression in E. Coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7122)
  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 7122)
  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_(+36GFP)-Cre
FEATURES             Location/Qualifiers
     source          1..7122
                     /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             997..2022
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="SNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLL
                     SVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLP
                     RPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFL
                     GIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVER
                     WISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQR
                     YLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLED
                     GD"
     CDS             2032..2049
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2068..2112
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(2150..2168)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2164..2211
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2248..2703
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2335..2354)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2545..2566
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(2818..3609,3610..3678))
                     /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     3441..3460
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3679..3771)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3852..4440
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4341..4360
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4594..4611
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4626..4768
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4694..4713)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4854..4876
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4870..5061)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5633..6715)
                     /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     6677..6696
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6716..6793)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6999..7018
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     join(7106..7122,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(7106..7122,1..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 aaactgagca aagatccgaa agaaaaacgc
      841 gatcacatgg ttctgttaga attcgtgacc gctgcaggca ttaagcacgg acgcgacgaa
      901 cgctacaaga ccggtggtag cggtggttct ggtggttctg gtggtagcgg cggtagcggt
      961 ggtagcggtg gtagcggtgg cagcggcggt accgcgagca atttactgac cgtacaccaa
     1021 aatttgcctg cattgccggt cgatgcaacg agtgatgagg ttcgcaagaa cctgatggac
     1081 atgttcaggg atcgccaggc gttttctgag catacctgga aaatgcttct gtccgtttgc
     1141 cggtcgtggg cggcatggtg caagttgaat aaccggaaat ggtttcccgc agaacctgaa
     1201 gatgttcgcg attatcttct atatcttcag gcgcgcggtc tggcagtaaa aactatccag
     1261 caacatttgg gccagctaaa catgcttcat cgtcggtccg ggctgccacg accaagtgac
     1321 agcaatgctg tttcactggt tatgcggcgt atccgaaaag aaaacgttga tgccggtgaa
     1381 cgtgcaaaac aggctctagc gttcgaacgc actgatttcg accaggttcg ttcactcatg
     1441 gaaaatagcg atcgctgcca ggatatacgt aatctggcat ttctggggat tgcttataac
     1501 accctgttac gtatagccga aattgccagg atcagggtta aagatatctc acgtactgac
     1561 ggtgggagaa tgttaatcca tattggcaga acgaaaacgc tggttagcac tgcaggtgta
     1621 gagaaggcac ttagcctggg ggtaactaaa ctggtcgagc gatggatttc cgtctctggt
     1681 gtagctgatg atccgaataa ctacctgttt tgccgggtca gaaaaaatgg tgttgccgcg
     1741 ccatctgcca ccagccagct atcaactcgc gccctggaag ggatttttga agcaactcat
     1801 cgattgattt acggcgctaa ggatgactct ggtcagagat acctggcctg gtctggacac
     1861 agtgcccgtg tcggagccgc gcgagatatg gcccgcgctg gagtttcaat accggagatc
     1921 atgcaagctg gtggctggac caatgtaaat attgtcatga actatatccg taacctggat
     1981 agtgaaacag gggcaatggt gcgcctgctg gaagatggcg acggcggatc ccatcaccac
     2041 caccatcact aatgactcga gtctggtaaa gaaaccgctg ctgcgaaatt tgaacgccag
     2101 cacatggact cgtctactag cgcagcttaa ttaacctagg ctgctgccac cgctgagcaa
     2161 taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga
     2221 ggaactatat ccggattggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg
     2281 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     2341 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     2401 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     2461 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     2521 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     2581 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     2641 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa
     2701 tttctggcgg cacgatggca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     2761 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     2821 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     2881 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     2941 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     3001 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     3061 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     3121 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     3181 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     3241 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     3301 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     3361 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     3421 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     3481 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     3541 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     3601 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     3661 gaaatgttga atactcatac tcttcctttt tcaatcatga ttgaagcatt tatcagggtt
     3721 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggtcatg
     3781 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     3841 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     3901 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     3961 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
     4021 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     4081 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     4141 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     4201 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     4261 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     4321 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     4381 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     4441 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     4501 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     4561 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     4621 gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatat
     4681 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct
     4741 atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc
     4801 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     4861 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag
     4921 ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg cgtccagctc
     4981 gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc
     5041 ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt
     5101 aatgataccg atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc
     5161 ccggttactg gaacgttgtg agggtaaaca actggcggta tggatgcggc gggaccagag
     5221 aaaaatcact cagggtcaat gccagcgctt cgttaataca gatgtaggtg ttccacaggg
     5281 tagccagcag catcctgcga tgcagatccg gaacataatg gtgcagggcg ctgacttccg
     5341 cgtttccaga ctttacgaaa cacggaaacc gaagaccatt catgttgttg ctcaggtcgc
     5401 agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta
     5461 accagtaagg caaccccgcc agcctagccg ggtcctcaac gacaggagca cgatcatgct
     5521 agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca agggcatcgg
     5581 tcgagatccc ggtgcctaat gagtgagcta acttacatta attgcgttgc gctcactgcc
     5641 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     5701 gagaggcggt ttgcgtattg ggcgccaggg tggtttttct tttcaccagt gagacgggca
     5761 acagctgatt gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg
     5821 tttgccccag caggcgaaaa tcctgtttga tggtggttaa cggcgggata taacatgagc
     5881 tgtcttcggt atcgtcgtat cccactaccg agatgtccgc accaacgcgc agcccggact
     5941 cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg
     6001 gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt
     6061 cgccttcccg ttccgctatc ggctgaattt gattgcgagt gagatattta tgccagccag
     6121 ccagacgcag acgcgccgag acagaactta atgggcccgc taacagcgcg atttgctggt
     6181 gacccaatgc gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa
     6241 tactgttgat gggtgtctgg tcagagacat caagaaataa cgccggaaca ttagtgcagg
     6301 cagcttccac agcaatggca tcctggtcat ccagcggata gttaatgatc agcccactga
     6361 cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta
     6421 ccatcgacac caccacgctg gcacccagtt gatcggcgcg agatttaatc gccgcgacaa
     6481 tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac gccaatcagc aacgactgtt
     6541 tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc atcgccgctt
     6601 ccactttttc ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg
     6661 tctgataaga gacaccggca tactctgcga catcgtataa cgttactggt ttcacattca
     6721 ccaccctgaa ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcgcc
     6781 attcgatggt gtccgggatc tcgacgctct cccttatgcg actcctgcat taggaagcag
     6841 cccagtagta ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag
     6901 atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg
     6961 ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc gatataggcg
     7021 ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc gtagaggatc
     7081 gagatcgatc tcgatcccgc gaaattaata cgactcacta ta
//
