LOCUS       sequence-59325-e        9123 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 9123)
  AUTHORS   Liu DS, Loh KH, Lam SS, White KA, Ting AY
  TITLE     Imaging trans-cellular neurexin-neuroligin interactions by enzymatic
            probe ligation.
  JOURNAL   PLoS One. 2013;8(2):e52823. doi: 10.1371/journal.pone.0052823. Epub 
            2013 Feb 14.
  PUBMED    23457442
REFERENCE   2  (bases 1 to 9123)
  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: "PLoS One.";
            date: "2013"; volume: "8(2):e52823. doi"; pages: " 
            10.1371/journal.pone.0052823. Epub 2013 Feb 14"
COMMENT     Sequence Label: pXL-CAG-3XAP-NLG1
FEATURES             Location/Qualifiers
     source          1..9123
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   746..808
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        1072..1375
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1378..1653
                     /label=chicken beta-actin promoter
     intron          1654..2663
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2587..2609
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2671..2690
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    2730..2754
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3009..3035
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3036..3080
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             3087..3131
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             3138..3182
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     regulatory      3656..3665
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(5620..5644)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(5735..5754)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5800..5855
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5854..5873)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     repeat_region   6449..6483
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(6974..6990)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6974..6990)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6987..7009)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6998..7014
                     /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(7022..7028,7029..7046,7047..7052))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7067..7088
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7205..7222)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7376..7964)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7456..7475)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8135..8926,8927..8995))
                     /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     8758..8777
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8996..9100)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcctcgtt cattcacgtt tttgaacccg tggaggacgg
      661 gcagactcgc ggtgcaaatg tgttttacag cgtgatggag cagatgaaga tgctcgacac
      721 gctgcagaac acgcagctag attaacccta gaaagataat catattgtga cgtacgttaa
      781 agataatcat gcgtaaaatt gacgcatgtg ttttatcggt ctgtatatcg aggtttattt
      841 attaatttga atagatatta agttttatta tatttacact tacatactaa taataaattc
      901 aacaaacaat ttatttatgt ttatttattt attaaaaaaa aacaaaaact caaaatttct
      961 tctataaagt aacaaaactt ttatcgaatt cctgcagccc gggggatcca ctagttatta
     1021 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
     1081 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     1141 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     1201 ctatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
     1261 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     1321 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatgggtcg
     1381 aggtgagccc cacgttctgc ttcactctcc ccatctcccc cccctcccca cccccaattt
     1441 tgtatttatt tattttttaa ttattttgtg cagcgatggg ggcggggggg gggggggcgc
     1501 gcgccaggcg gggcggggcg gggcgagggg cggggcgggg cgaggcggag aggtgcggcg
     1561 gcagccaatc agagcggcgc gctccgaaag tttcctttta tggcgaggcg gcggcggcgg
     1621 cggccctata aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgttgcc ttcgccccgt
     1681 gccccgctcc gcgccgcctc gcgccgcccg ccccggctct gactgaccgc gttactccca
     1741 caggtgagcg ggcgggacgg cccttctcct ccgggctgta attagcgctt ggtttaatga
     1801 cggctcgttt cttttctgtg gctgcgtgaa agccttaaag ggctccggga gggccctttg
     1861 tgcggggggg agcggctcgg ggggtgcgtg cgtgtgtgtg tgcgtgggga gcgccgcgtg
     1921 cggcccgcgc tgcccggcgg ctgtgagcgc tgcgggcgcg gcgcggggct ttgtgcgctc
     1981 cgcgtgtgcg cgaggggagc gcggccgggg gcggtgcccc gcggtgcggg ggggctgcga
     2041 ggggaacaaa ggctgcgtgc ggggtgtgtg cgtggggggg tgagcagggg gtgtgggcgc
     2101 ggcggtcggg ctgtaacccc cccctgcacc cccctccccg agttgctgag cacggcccgg
     2161 cttcgggtgc ggggctccgt gcggggcgtg gcgcggggct cgccgtgccg ggcggggggt
     2221 ggcggcaggt gggggtgccg ggcggggcgg ggccgcctcg ggccggggag ggctcggggg
     2281 aggggcgcgg cggccccgga gcgccggcgg ctgtcgaggc gcggcgagcc gcagccattg
     2341 ccttttatgg taatcgtgcg agagggcgca gggacttcct ttgtcccaaa tctggcggag
     2401 ccgaaatctg ggaggcgccg ccgcaccccc tctagcgggc gcgggcgaag cggtgcggcg
     2461 ccggcaggaa ggaaatgggc ggggagggcc ttcgtgcgtc gccgcgccgc cgtccccttc
     2521 tccctctcca gcctcggggc tgtccgcggg gggacggctg ccttcggggg ggacggggca
     2581 gggcggggtt cggcttctgg cgtgtgaccg gcggctctag agcctctgct aaccatgttc
     2641 atgccttctt ctttttccta cagctcctgg gcaacgtgct ggttattgtg ctgtctcatc
     2701 attttggcaa agaattcctc gaggatatca caagtttgta caaaaaagca ggctccgaat
     2761 tcgcccttcg taactataac ggtcctaagg tagcgaatag tgaaccgtca gatccgctag
     2821 cgctaccgga ctcagatctc gagctcaagc ttgggaccat ggcacttccc agatgcatgt
     2881 ggccaaatta tgtttggaga gctatgatgg catgtgtggt ccacagggga tccggtgccc
     2941 cattgactct ctgcttgttg ggatgtttgc tacagacttt tcacgtactc tctcaaaagt
     3001 atcgatcgta tccatatgat gttccagatt atgctggcct gaacgacatc ttcgaggccc
     3061 agaagatcga gtggcacgag ggcagcggac tgaatgatat ttttgaagct caaaagattg
     3121 aatggcatga aggaggaggg ctgaacgata tctttgaggc acagaaaatc gaatggcatg
     3181 agggaggcgg agtcgacgca ttggatgatg tagacccatt ggttactact aactttggca
     3241 agattagggg aattaagaaa gaactcaata atgaaatttt gggtcctgtc attcagtttc
     3301 ttggggttcc atatgccgct ccaccaacag gagaacatcg tttccagcct ccagaaccac
     3361 catctccctg gtctgacatc cggaacgcca ctcagtttgc tcctgtatgt ccccagaata
     3421 tcattgatgg cagattgcct gaagttatgc ttcctgtgtg gttcactaat aacttggatg
     3481 tggtttcatc atacgtccaa gaccagagtg aagactgtct atacttaaac atctatgtcc
     3541 caactgaaga tggtcccctt acaaagaaac acacagatga tttaggtgat aatgacggtg
     3601 ctgaagatga agatattcgg gacagtgggg gtcccaaacc agtgatggtg tacatccatg
     3661 gcggctctta catggaaggt actggaaatc tgtatgatgg gagtgtcttg gcaagctatg
     3721 gcaatgtgat cgtcatcaca gtcaactatc ggcttggggt acttggcttc ttgagcacag
     3781 gggatcaggc tgccaaagga aactacgggc tccttgacct catccaggcc ctaagatgga
     3841 ccagcgagaa cattgggttc tttggtggtg accccttgcg aatcactgtg tttggatcag
     3901 gcgctggggg ttcatgtgtc aacctgctga ctttatccca ttattctgaa ggtaaccgtt
     3961 ggagcaattc aaccaaagga ctttttcaac gagcaatagc tcagagtgga acagcccttt
     4021 ccagctgggc tgttagtttc cagcctgcaa aatacgctag aattctggcc acaaaagttg
     4081 gctgcaatgt ttcagataca gtagagttag tagaatgcct gcagaagaag ccttacaaag
     4141 aacttgttga tcaagatgtt caaccagccc gataccacat agcctttgga cctgtgatcg
     4201 atggtgatgt aataccagat gaccctcaga tactgatgga acaaggagag ttcctcaact
     4261 atgatataat gttgggagtt aaccaagggg aagggttgaa gtttgtcgaa aatatagtag
     4321 atagtgatga tggtgtatca gccagtgatt tcgactttgc tgtttctaat tttgttgata
     4381 atttatatgg atatcctgaa ggcaaagatg ttttgagaga aaccattaaa ttcatgtata
     4441 ctgactgggc tgatcgccat aatcctgaaa ctagaaggaa gacattgttg gctttgttta
     4501 cggaccatca atgggtagca cctgctgtgg ccacagcaga ccttcactcg aactttggct
     4561 cacctacata cttctatgcc ttttatcatc attgccaaac agatcaagtt ccagcttggg
     4621 ctgatgcagc tcatggggat gaggttccct atgtgttggg aatccccatg attggcccta
     4681 cagagttatt tccttgcaat ttctccaaga atgatgtgat gttgagtgca gtagtaatga
     4741 catactggac gaattttgct aaaactggtg acccaaatca accagttcct caagacacaa
     4801 aattcatcca taccaaaccc aaccgctttg aagaagtagc atggaccaga tattcccaga
     4861 aagaccagct ttatctccat attggattaa aaccgagagt taaagagcat tacagagcca
     4921 ataaggtaaa tctctggctg gagctggtac ctcatctgca taatctcaat gacatttctc
     4981 agtatacctc gacaacaact aaagtgccat ccacggacat cactctcaga cctacaagga
     5041 agaattccac accagtcaca tcagcctttc ccactgccaa acaggatgat cccaagcaac
     5101 aaccaagccc cttctcggtg gatcagaggg actactccac agagctaagt gtcactatcg
     5161 cagtgggggc ctctctgctg tttctcaaca tcttggcttt tgcagccctg tactacaaga
     5221 aggataagag gagacatgat gtccaccgga ggtgcagccc tcagcgcacg accaccaacg
     5281 acctaaccca tgctccagaa gaggaaatta tgtctctcca aatgaagcac actgacttgg
     5341 atcacgagtg tgagtccatc catccacatg aggtggttct tcggaccgcc tgtcccccag
     5401 attatactct agctatgagg aggtcacctg atgatattcc actaatgaca cctaacacca
     5461 tcacaatgat tcccaacact ataccaggga ttcagccctt acatacattc aacacattta
     5521 ctggaggaca gaataataca ctgccccatc cccacccaca cccccattca cattcaacaa
     5581 ccagggtata gaagctagcg ggggccaagg gcgaattcga cccagctttc ttgtacaaag
     5641 tggtgattcg cggccgcact cctcaggtgc aggctgccta tcagaaggtg gtggctggtg
     5701 tggccaatgc cctggctcac aaataccact gagatctttt tccctctgcc aaaaattatg
     5761 gggacatcat gaagcccctt gagcatctga cttctggcta ataaaggaaa tttattttca
     5821 ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg aaggacatat gggagggcaa
     5881 atcatttaaa acatcagaat gagtatttgg tttagagttt ggcaacatat gcccatatgc
     5941 tggctgccat gaacaaaggt tggctataag aggtcatcag tatatgaaac agcccctgct
     6001 gtccattcta tccatagaaa gctgactgag tagatttttt tatatttgtt tgtgtatttt
     6061 tctacatccc taaatttcna catgtttact agcgaatttt cctccctctg actactccca
     6121 gtcatagctg tccctcttct cttatgaaga tccctcgacc tgcagcccaa gcttgatatc
     6181 tataacaaga aaatatatat ataataagtt atcacgtaag tagaacatga aataacaata
     6241 taattatcgt atgagttaaa tcttaaaagt cacgtaaaag ataatcatgc gtcattttga
     6301 ctcacgcggt cgttatagtt caaaatcagt gacacttacc gcattgacaa gcacgcctca
     6361 cgggagctcc aagcggcgac tgagatgtcc taaatgcaca gcgacggatt cgcgctattt
     6421 agaaagagag agcaatattt caagaatgca tgcgtcaatt ttacgcagac tatctttcta
     6481 gggttaatct agctgcatca ggatcatatc gtcgggtctt ttttccggct cagtcatcgc
     6541 ccaagctggc gctatctggg catcggggag gaagaagccc gtgccttttc ccgcgaggtt
     6601 gaagcggcat ggaaagagtt tgccgaggat gactgctgct gcattgacgt tgagcgaaaa
     6661 cgcacgttta ccatgatgat tcgggaaggt gtggccatgc acgcctttaa cggtgaactg
     6721 ttcgttcagg ccacctggga taccagttcg tcgcggcttt tccggacaca gttccggatg
     6781 gtcagcccga agcgcatcag caacccgaac aataccggcg acagccggaa ctgccgtgcc
     6841 ggtgtgcaga ttaatgacag cggtgcggcg ctgggatatt acgtcagcga ggacgggtat
     6901 cctggctgga tgccgcagaa atggacatgg ataccccgtg agttacccgg cgggcgcgct
     6961 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     7021 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     7081 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     7141 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     7201 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     7261 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     7321 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     7381 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     7441 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     7501 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     7561 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     7621 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     7681 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     7741 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     7801 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     7861 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     7921 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     7981 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     8041 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     8101 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     8161 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     8221 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     8281 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     8341 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     8401 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     8461 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     8521 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     8581 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     8641 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     8701 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     8761 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     8821 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     8881 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     8941 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     9001 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     9061 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     9121 cac
//
