LOCUS       sequence-59323-e        8624 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 8624)
  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 8624)
  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-mBirA272-NRX3b D137A
FEATURES             Location/Qualifiers
     source          1..8624
                     /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             3774..4544
                     /codon_start=1
                     /product="N-terminally truncated mutant of the E. coli 
                     biotin ligase BioID engineered for improved catalytic 
                     efficiency (Branon et al., 2018)"
                     /label=miniTurbo
                     /translation="IQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSGDACI
                     AEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAIGLSLVIGIVMAEVLRKLGADK
                     VRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQGWITLQ
                     EAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIGDKEIF
                     GISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             4722..4769
                     /codon_start=1
                     /product="two tandem FLAG(R) epitope tags"
                     /label=2xFLAG
                     /translation="DYKDDDDKDYKDDDDK"
     protein_bind    complement(5092..5116)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(5207..5226)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5272..5327
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5326..5345)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     repeat_region   5950..5984
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(6475..6491)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6475..6491)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6488..6510)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6499..6515
                     /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(6523..6529,6530..6547,6548..6553))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6568..6589
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6706..6723)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6877..7465)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6957..6976)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7636..8427,8428..8496))
                     /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"
     promoter        complement(8497..8601)
                     /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 ggcttcgaag
     2761 gagatagaac catgcacctg agaatccacg cgagacggag ccctcctcgc cggccggcct
     2821 ggacgcttgg gatctggttc ctgttctggg gatgtatcgt cagctctgta tggagttctt
     2881 ctaatgtagc ttcctcctcc tccacctctt cctcgccggg gtctcactct cagcacgagc
     2941 accatttcca tggcagcaag catcactcag tgcctatttc tatctatcgt tcccctgttt
     3001 cccttcgagg aggacacgct ggcgctacgt acatctttgg gaaaagtggt gggcttatcc
     3061 tctacacctg gccagccaat gacaggccca gcacgcggtc tgaccgcctt gccgtgggct
     3121 tcagcaccac tgtgaaggat ggcatcttgg tccgcatcga cagtgctcca ggacttggtg
     3181 ccttcctcca gcttcacata gaacagggga aaattggagt tgtcttcaac attggcacag
     3241 ttgacatctc catcaaagag gagagaaccc ctgtaaatga cggcaaatac catgtggtac
     3301 gcttcaccag gaacggcggc aacgccaccc tgcaggtgga caactggcca gtgaatgaac
     3361 attatcctac aggccggcag ttaaccatct tcaacactca ggcgcaaata gccattggtg
     3421 gaaaggacaa aggacgcctc ttccaaggcc aactctctgg gctctattat gatggtttga
     3481 aagtactgaa catggcggct gagaacaacc ccaatattaa aatcaatgga agtgttcggc
     3541 tggttggaga agtcccatca attttgggaa caacacagac gaccatgaag gataacaccg
     3601 tgcccctgaa actgattgct ctgctggcta atggcgagtt ccatagtggc gaacagctgg
     3661 gagaaaccct gggcatgtcc agggccgcta tcaacaagca cattcagact ctgcgcgact
     3721 ggggcgtgga cgtgttcacc gtgcccggaa agggctactc tctgcccgag cctatccagc
     3781 tgctgaacgc taaacagatt ctgggacagc tggacggcgg gagcgtggca gtcctgcctg
     3841 tgatcgactc caccaatcag tacctgctgg atcgaatcgg cgagctgaag agtggggatg
     3901 cttgcattgc agaatatcag caggcaggaa gaggacggag agggaggaaa tggttctctc
     3961 cttttggagc taacctgtac ctgagtatgt tttggcgcct ggagcaggga ccagcagcaa
     4021 tcggcctgag cctggtcatc ggaattgtca tggcagaagt gctgcgaaag ctgggagcag
     4081 acaaggtgcg agtcaaatgg cccaatgacc tgtatctgca ggatagaaag ctggcaggca
     4141 tcctggtgga gctgaccgga aaaacaggcg atgctgcaca gatcgtcatt ggcgccggga
     4201 ttaacatggc tatgaggcgc gtggaggaaa gcgtggtcaa tcagggctgg atcacactgc
     4261 aggaagcagg gattaacctg gacaggaata ctctggccgc tatgctgatc cgagagctgc
     4321 gggcagccct ggaactgttc gagcaggaag gcctggctcc atatctgtca cggtgggaga
     4381 agctggataa cttcatcaat agacccgtga agctgatcat tggggacaaa gagattttcg
     4441 ggattagccg gggaattgat aaacagggag ccctgctgct ggaacaggac ggaattatca
     4501 aaccctggat gggcggagaa atcagtctgc ggtctgccga aaagtccatg ccaccagaaa
     4561 tgtctactac tgtcatggaa accactacta caatggcgac taccacaacc cgtaagaatc
     4621 gctctacagc cagcattcag ccaacatcag atgatcttgt ttcatctgct gaatgttcaa
     4681 gtgatgatga agactttgtt gaatgtgagc cgagtacagc agactacaag gatgacgacg
     4741 ataaggatta caaagacgat gatgacaaaa accccacgga gccgggaatc agacgggttc
     4801 cgggggcctc agaggtgatc cgggagtcga gcagcacaac agggatggtc gtcggcattg
     4861 tggctgctgc cgccctctgc atcttgatcc tcctgtacgc catgtacaag tacaggaaca
     4921 gggacgaggg gtcctatcaa gtggacgaga cgcggaacta catcagcaac tccgcccaga
     4981 gcaacggcac gctcatgaag gagaagcagc agagctcgaa gagcggccac aagaaacaga
     5041 aaaacaagga cagggagtat tacgtgttct gctacgagaa cgaggtgtag gacccagctt
     5101 tcttgtacaa agtggtgatt cgcggccgca ctcctcaggt gcaggctgcc tatcagaagg
     5161 tggtggctgg tgtggccaat gccctggctc acaaatacca ctgagatctt tttccctctg
     5221 ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc taataaagga
     5281 aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat
     5341 atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt ttggcaacat
     5401 atgcccatat gctggctgcc atgaacaaag gttggctata aagaggtcat cagtatatga
     5461 aacagccccc tgctgtccat tccttattcc atagaaaagc cttgacttga ggttagattt
     5521 tttttatatt ttgttttgtg ttattttttt ctttaacatc cctaaaattt tccttacatg
     5581 ttttactagc cagatttttc ctcctctcct gactactccc agtcatagct gtccctcttc
     5641 tcttatgaag atccctcgac ctgcagccca agcttgatat ctataacaag aaaatatata
     5701 tataataagt tatcacgtaa gtagaacatg aaataacaat ataattatcg tatgagttaa
     5761 atcttaaaag tcacgtaaaa gataatcatg cgtcattttg actcacgcgg tcgttatagt
     5821 tcaaaatcag tgacacttac cgcattgaca agcacgcctc acgggagctc caagcggcga
     5881 ctgagatgtc ctaaatgcac agcgacggat tcgcgctatt tagaaagaga gagcaatatt
     5941 tcaagaatgc atgcgtcaat tttacgcaga ctatctttct agggttaatc tagctgcatc
     6001 aggatcatat cgtcgggtct tttttccggc tcagtcatcg cccaagctgg cgctatctgg
     6061 gcatcgggga ggaagaagcc cgtgcctttt cccgcgaggt tgaagcggca tggaaagagt
     6121 ttgccgagga tgactgctgc tgcattgacg ttgagcgaaa acgcacgttt accatgatga
     6181 ttcgggaagg tgtggccatg cacgccttta acggtgaact gttcgttcag gccacctggg
     6241 ataccagttc gtcgcggctt ttccggacac agttccggat ggtcagcccg aagcgcatca
     6301 gcaacccgaa caataccggc gacagccgga actgccgtgc cggtgtgcag attaatgaca
     6361 gcggtgcggc gctgggatat tacgtcagcg aggacgggta tcctggctgg atgccgcaga
     6421 aatggacatg gataccccgt gagttacccg gcgggcgcgc ttggcgtaat catggtcata
     6481 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     6541 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     6601 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     6661 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     6721 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     6781 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     6841 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     6901 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     6961 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     7021 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     7081 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     7141 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     7201 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     7261 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     7321 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     7381 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     7441 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     7501 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     7561 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     7621 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     7681 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     7741 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     7801 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     7861 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     7921 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     7981 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     8041 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     8101 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     8161 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     8221 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     8281 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     8341 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     8401 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     8461 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     8521 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     8581 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccac
//
