LOCUS       sequence-316658-        9443 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9443)
  AUTHORS   Kallay LM, McNickle A, Brennwald PJ, Hubbard AL, Braiterman LT
  TITLE     Scribble associates with two polarity proteins, Lgl2 and Vangl2, via
            distinct molecular domains.
  JOURNAL   J Cell Biochem. 2006 Oct 1;99(2):647-64.
  PUBMED    16791850
REFERENCE   2  (bases 1 to 9443)
  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: "J Cell 
            Biochem."; date: "2006-10-1"; volume: "99(2)"; pages: "647-64"
COMMENT     Sequence Label: pLB1080
FEATURES             Location/Qualifiers
     source          1..9443
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      646..655
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(652..654,655..657,658..1368)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(697..718)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(958..977)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1305..1326
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5961..6095
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5998..6017)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6052..6071
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    6104..6137
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        complement(6149..6167)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(6150..6167)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     repeat_region   6240..6342
                     /label=ITR
                     /note="inverted terminal repeat of human adenovirus 
                     serotype 5"
     rep_origin      6426..6881
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6513..6532)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6723..6744
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6907..7011
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7012..7080,7081..7872)
                     /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     complement(7230..7249)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8043..8631
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8532..8551
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8785..8802
                     /label=L4440
                     /note="L4440 vector, forward primer"
     repeat_region   8881..8983
                     /label=ITR
                     /note="inverted terminal repeat of human adenovirus 
                     serotype 5"
     misc_signal     9071..9221
                     /label=Ad5 Psi
                     /note="packaging signal for adenovirus serotype 5"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgta
      601 agcttgcatg cctgcaggtc gactctagct agaggatccc cggtcgccac catggtgagc
      661 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
      721 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
      781 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
      841 accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac
      901 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
      961 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     1021 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     1081 tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag
     1141 gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac
     1201 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     1261 acccagtcca agctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     1321 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagtc cggactcaga
     1381 tccatgcctg aacaggagag acagatcaca gccagagaag gggccagtcg gaaaatctta
     1441 tctaagcttt ctttgcctac ccgtgcctgg gaaccagcaa tgaagaagag ttttgctttt
     1501 gacaatgttg gctatgaagg tggtctggat ggcctgggcc cttcttctca ggtggccacc
     1561 agcacagtca ggatcttggg catgacttgc cagtcatgtg tgaagtccat tgaggacagg
     1621 atttccaatt tgaaaggcat catcagcatg aaggtttccc tggaacaagg cagtgccact
     1681 gtgaaatatg tgccatcggt tgtgtgcctg caacaggttt gccatcaaat tggggacatg
     1741 ggcttcgagg ccagcattgc agaaggaaag gcagcctcct ggccctcaag gtccttgcct
     1801 gcccaggagg ctgtggtcaa gctccgggtg gagggcatga cctgccagtc ctgtgtcagc
     1861 tccattgaag gcaaggtccg gaaactgcaa ggagtagtga gagtcaaagt ctcactcagc
     1921 aaccaagagg ccgtcatcac ttatcagcct tatctcattc agcccgaaga cctcagggac
     1981 catgtaaatg acatgggatt tgaagctgcc atcaagagca aagtggctcc cttaagcctg
     2041 ggaccaattg atattgagcg gttacaaagc actaacccaa agagaccttt atcttctgct
     2101 aaccagaatt ttaataattc tgagaccttg gggcaccaag gaagccatgt ggtcaccctc
     2161 caactgagaa tagatggaat gcattgtaag tcttgcgtct tgaatattga agaaaatatt
     2221 ggccagctcc taggggttca aagtattcaa gtctccttgg agaacaaaac tgcccaagta
     2281 aagtatgacc cttcttgtac cagcccagtg gctctgcaga gggctatcga ggcacttcca
     2341 cctgggaatt ttaaagtttc tcttcctgat ggagccgaag ggagtgggac agatcacagg
     2401 tcttccagtt ctcattcccc tggctcccca ccgagaaacc aggtccaggg cacatgcagt
     2461 accactctga ttgccattgc cggcatgacc tgtgcatcct gtgtccattc cattgaaggc
     2521 atgatctccc aactggaagg ggtgcagcaa atatcggtgt ctttggccga agggactgca
     2581 acagttcttt ataatcccgc tgtaattagc ccagaagaac tcagagctgc tatagaagac
     2641 atgggatttg aggcttcagt cgtttctgaa agctgttcta ctaaccctct tggaaaccac
     2701 agtgctggga attccatggt gcaaactaca gatggtacac ctacatctct gcaggaagtg
     2761 gctccccaca ctgggaggct ccctgcaaac catgccccgg acatcttggc aaagtcccca
     2821 caatcaacca gagcagtggc accgcagaag tgcttcttac agatcaaagg catgacctgt
     2881 gcatcctgtg tgtctaacat agaaaggaat ctgcagaaag aagctggtgt tctctccgtg
     2941 ttggttgcct tgatggcagg aaaggcagag atcaagtatg acccagaggt catccagccc
     3001 ctcgagatag ctcagttcat ccaggacctg ggttttgagg cagcagtcat ggaggactac
     3061 gcaggctccg atggcaacat tgagctgaca atcacaggga tgacctgcgc gtcctgtgtc
     3121 cacaacatag agtccaaact cacgaggaca aatggcatca cttatgcctc cgttgccctt
     3181 gccaccagca aagcccttgt taagtttgac ccggaaatta tcggtccacg ggatattatc
     3241 aaaattattg aggaaattgg ctttcatgct tccctggccc agagaaaccc caacgctcat
     3301 cacttggacc acaagatgga aataaagcag tggaagaagt ctttcctgtg cagcctggtg
     3361 tttggcatcc ctgtcatggc cttaatgatc tatatgctga tacccagcaa cgagccccac
     3421 cagtccatgg tcctggacca caacatcatt ccaggactgt ccattctaaa tctcatcttc
     3481 tttatcttgt gtacctttgt ccagctcctc ggtgggtggt acttctacgt tcaggcctac
     3541 aaatctctga gacacaggtc agccaacatg gacgtgctca tcgtcctggc cacaagcatt
     3601 gcttatgttt attctctggt catcctggtg gttgctgtgg ctgagaaggc ggagaggagc
     3661 cctgtgacat tcttcgacac gccccccatg ctctttgtgt tcattgccct gggccggtgg
     3721 ctggaacact tggcaaagag caaaacctca gaagccctgg ctaaactcat gtctctccaa
     3781 gccacagaag ccaccgttgt gacccttggt gaggacaatt taatcatcag ggaggagcaa
     3841 gtccccatgg agctggtgca gcggggcgat atcgtcaagg tggtccctgg gggaaagttt
     3901 ccagtggatg ggaaagtcct ggaaggcaat accatggctg atgagtccct catcacagga
     3961 gaagccatgc cagtcactaa gaaacccgga agcactgtaa ttgcggggtc tataaatgca
     4021 catggctctg tgctcattaa agctacccac gtgggcaatg acaccacttt ggctcagatt
     4081 gtgaaactgg tggaagaggc tcagatgtca aaggcaccca ttcagcagct ggctgaccgg
     4141 tttagtggat attttgtccc atttatcatc atcatgtcaa ctttgacgtt ggtggtatgg
     4201 attgtaatcg gttttatcga ttttggtgtt gttcagaaat actttcctaa ccccaacaag
     4261 cacatctccc agacagaggt gatcatccgg tttgctttcc agacgtccat cacggtgctg
     4321 tgcattgcct gcccctgctc cctggggctg gccacgccca cggctgtcat ggtgggcacc
     4381 ggggtggccg cgcagaacgg catcctcatc aagggaggca agcccctgga gatggcgcac
     4441 aagataaaga ctgtgatgtt tgacaagact ggcaccatta cccatggcgt ccccagggtc
     4501 atgcgggtgc tcctgctggg ggatgtggcc acactgcccc tcaggaaggt tctggctgtg
     4561 gtggggactg cggaggccag cagtgaacac cccttgggcg tggcagtcac caaatactgt
     4621 aaagaggaac ttggaacaga gaccttggga tactgcacgg acttccaggc agtgccaggc
     4681 tgtggaattg ggtgcaaagt cagcaacgtg gaaggcatcc tggcccacag tgagcgccct
     4741 ttgagtgcac cggccagtca cctgaatgag gctggcagcc ttcccgcaga aaaagatgca
     4801 gccccccaga ccttctctgt gctgattgga aaccgtgagt ggctgaggcg caacggttta
     4861 accatttcta gcgatgtcag tgacgctatg acagaccacg agatgaaagg acagacagcc
     4921 atcctggtgg ctattgacgg tgtgctctgt gggatgatcg caatcgcaga cgctgtcaag
     4981 caggaggctg ccctggctgt gcacacgctg cagagcatgg gtgtggacgt ggttctgatc
     5041 acgggggaca accggaagac agccagagct attgccaccc aggttggcat caacaaagtc
     5101 tttgcagagg tgctgccttc gcacaaggtg gccaaggtcc aggagctcca gaataaaggg
     5161 aagaaagtcg ccatggtggg ggatggggtc aatgactccc cggccttggc ccaggcagac
     5221 atgggtgtgg ccattggcac cggcacggat gtggccatcg aggcagccga cgtcgtcctt
     5281 atcagaaatg atttgctgga tgtggtggct agcattcacc tttccaagag gactgtccga
     5341 aggatacgca tcaacctggt cctggcactg atttataacc tggttgggat acccattgca
     5401 gcaggtgtct tcatgcccat cggcattgtg ctgcagccct ggatgggctc agcggccatg
     5461 gcagcctcct ctgtgtctgt ggtgctctca tccctgcagc tcaagtgcta taagaagcct
     5521 gacctggaga ggtatgaggc acaggcgcat ggccacatga agcccctgac ggcatcccag
     5581 gtcagtgtgc acataggcat ggatgacagg tggcgggact cccccagggc cacaccatgg
     5641 gaccaggtca gctatgtcag ccaggtgtcg ctgtcctccc tgacgtccga caagccatct
     5701 cggcacagcg ctgcagcaga cgatgatggg gacaagtggt ctctgctcct gaatggcagg
     5761 gatgaggagc agtacatctg atgacttcag gcaggcgggc cggggcaggg acttgcctcc
     5821 actcaccata agctgagcag gacagccgga tccactagtc cagtgtggtg gaattctttg
     5881 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     5941 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     6001 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     6061 ccaaactcat caatgtatct tatcatgtct ggatcatcga tccataactt cgtataatgt
     6121 atgctatacg aagttatcca gatctggttc tatagtgtca cctaaatcgt atgtgtatga
     6181 tacataaggt tatgtattaa ttgtagccgc gttctaacga caatatgtcc atagggcccc
     6241 tacgtcaccc gccccgttcc cacgccccgc gccacgtcac aaactccacc ccctcattat
     6301 catattggct tcaatccaaa ataaggtata ttattgatga tggccgcagc ggccctggcg
     6361 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     6421 atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     6481 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     6541 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     6601 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     6661 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     6721 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     6781 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     6841 atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga
     6901 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     6961 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     7021 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     7081 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     7141 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     7201 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     7261 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     7321 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     7381 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     7441 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     7501 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     7561 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     7621 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     7681 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     7741 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     7801 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     7861 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     7921 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     7981 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     8041 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     8101 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     8161 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     8221 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     8281 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     8341 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     8401 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     8461 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     8521 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     8581 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     8641 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     8701 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     8761 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     8821 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagggg ccgctgcggc
     8881 catcatcaat aatatacctt attttggatt gaagccaata tgataatgag ggggtggagt
     8941 ttgtgacgtg gcgcggggcg tgggaacggg gcgggtgacg tagtagtgtg gcggaagtgt
     9001 gatgttgcaa gtgtggcgga acacatgtaa gcgacggatg tggcaaaagt gacgtttttg
     9061 gtgtgcgccg gtgtacacag gaagtgacaa ttttcgcgcg gttttaggcg gatgttgtag
     9121 taaatttggg cgtaaccgag taagatttgg ccattttcgc gggaaaactg aataagagga
     9181 agtgaaatct gaataatttt gtgttactca tagcgcgtaa tatttgtcta gggccgcggg
     9241 gactttgacc gtttacgtgg agactcgccc aggtgttttt ctcaggtgtt ttccgcgttc
     9301 cgggtcaaag ttggcgtttt attattatag tcagctgacg tgtagtgtat ttatacccgg
     9361 tgagttcctc aagaggccac tcttgagtgc cagcgagtag agttttctcc tccgagccgc
     9421 tccgacaccg ggactcgagt gtt
//
