LOCUS       sequence-7782-ex        5606 bp    DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5606)
  AUTHORS   Bateman JR, Lee AM, Wu CT
  TITLE     Site-specific transformation of Drosophila via phiC31 
            integrase-mediated cassette exchange.
  JOURNAL   Genetics. 2006 Jun . 173(2):769-77.
  PUBMED    16547094
REFERENCE   2  (bases 1 to 5606)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Genetics. 
            2006 Jun . 173(2):769-77."
COMMENT     Sequence Label: piB-GFP
FEATURES             Location/Qualifiers
     source          1..5606
                     /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"
     primer_bind     623..642
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        623..641
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    686..755
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     primer_bind     945..961
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     946..962
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1014..1252
                     /label=hsp70 promoter
                     /note="Drosophila melanogaster hsp70Bb promoter"
     primer_bind     1199..1220
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     intron          2101..2166
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             2296..2316
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2741..2875
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2778..2797)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2832..2851
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3043..3059)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3043..3059)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    3256..3325
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     promoter        complement(3418..3436)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3457..3473)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3457..3473)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3470..3492)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3481..3497
                     /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(3505..3511,3512..3529,3530..3535))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3550..3571
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3688..3705)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3859..4447)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3939..3958)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4618..5409,5410..5478))
                     /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     5241..5260
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5479..5583)
                     /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 gccagtgaat tgtaatacga ctcactatag ggcgaattgg gtaccgggcc
      661 ccccctcgac gatgtaggtc acggtctcga agccgcggtg cgggtgccag ggcgtgccct
      721 tgggctcccc gggcgcgtac tccacctcac ccatctggtc catcatgatg aacgggtcga
      781 ggtggcggta gttgatcccg gcgaacgcgc ggcgcaccgg gaagccctcg ccctcgaaac
      841 cgctgggcgc ggtggtcacg gtgagcacgg gacgtgcgac ggcgtcggcg ggtgcggata
      901 cgcggggcag cgtcagcggg ttctcgacgg tcacggcggg catgtcgagg tcgacggtat
      961 cgataagctt gatatcgaat tcctgcagcc cgggggatcc ggcgaaaaga gcgccggagt
     1021 ataaatagag gcgcttcgtc tacggagcga caattcaatt caaacaagca aagtgaacac
     1081 gtcgctaagc gaaagctaag caaataaaca agcgcagctg aacaagctaa acaatctgca
     1141 gtaaagtgca agttaaagtg aatcaattaa aagtaaccag caaccaagta aatcaactgc
     1201 aactactgaa atctgccaag aagtaattat tgaatacaag aagagaactc tgaatacttt
     1261 caacaagtta ccgagaaaga agaactcaca cacaatgcct actagtaagg gcgaggaact
     1321 gttcactggc gtggtcccaa ttctcgtgga actggatggc gatgtgaatg ggcacaaatt
     1381 ttctgtcagc ggagagggtg aaggtgatgc cacatacgga aagctcaccc tgaaattcat
     1441 ctgcaccact ggaaagctcc ctgtgccatg gccaacactg gtcactacct tcacctatgg
     1501 cgtgcagtgc ttttccagat acccagacca tatgaagcag catgactttt tcaagagcgc
     1561 catgcccgag ggctatgtgc aggagagaac catctttttc aaagatgacg ggaactacaa
     1621 gacccgcgct gaagtcaagt tcgaaggtga caccctggtg aatagaatcg agctgaaggg
     1681 cattgacttt aaggaggatg gaaacattct cggccacaag ctggaataca actataactc
     1741 ccacaatgtg tacatcatgg ccgacaagca aaagaatggc atcaaggtca acttcaagat
     1801 cagacacaac attgaggatg gctctgtgca gctggccgac cattatcaac agaacactcc
     1861 aatcggcgac ggccctgtgc tcctcccaga caaccattac ctgtccaccc agtctgccct
     1921 gtctaaagat cccaacgaaa agagagacca catggtcctg ctggagtttg tgaccgctgc
     1981 tgggatcaca catggcatgg acgagctgta caagtgatct agaggatctt tgtgaaggaa
     2041 ccttacttct gtggtgtgac ataattggac aaactaccta cagagattta aagctctaag
     2101 gtaaatataa aatttttaag tgtataatgt gttaaactac tgattctaat tgtttgtgta
     2161 ttttagattc caacctatgg aactgatgaa tgggagcagt ggtggaatgc ctttaatgag
     2221 gaaaacctgt tttgctcaga agaaatgcca tctagtgatg atgaggctac tgctgactct
     2281 caacattcta ctcctccaaa aaagaagaga aaggtagaag accccaagga ctttccttca
     2341 gaattgctaa gttttttgag tcatgctgtg tttagtaata gaactcttgc ttgctttgct
     2401 atttacacca caaaggaaaa agctgcactg ctatacaaga aaattatgga aaaatattct
     2461 gtaaccttta taagtaggca taacagttat aatcataaca tactgttttt tcttactcca
     2521 cacaggcata gagtgtctgc tattaataac tatgctcaaa aattgtgtac ctttagcttt
     2581 ttaatttgta aaggggttaa taaggaatat ttgatgtata gtgccttgac tagagatcat
     2641 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     2701 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     2761 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     2821 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatcgggcg
     2881 agctcgaatt ggtcgacctg cagccaagct ttgcgtactc gcaaattatt aaaaataaaa
     2941 ctttaaaaat aatttcgtct aattaatatt atgagttaat tcaaacccca cggacatgct
     3001 aaggaagggc gaattccagc acactggcgg ccgttactag tggatccact agttctagag
     3061 cggccgcatg cccgccgtga ccgtcgagaa cccgctgacg ctgccccgcg tatccgcacc
     3121 cgccgacgcc gtcgcacgtc ccgtgctcac cgtgaccacc gcgcccagcg gtttcgaggg
     3181 cgagggcttc ccggtgcgcc gcgcgttcgc cgggatcaac taccgccacc tcgacccgtt
     3241 catcatgatg gaccagatgg gtgaggtgga gtacgcgccc ggggagccca agggcacgcc
     3301 ctggcacccg caccgcggct tcgagaccga cctacatcac ggccgaatga agggcgaatt
     3361 ctgcagatat ccatcacact ggcggccgcc accgcggtgg agctccagct tttgttccct
     3421 ttagtgaggg ttaatttcga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     3481 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     3541 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     3601 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     3661 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     3721 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     3781 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     3841 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3901 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3961 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     4021 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     4081 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     4141 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     4201 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     4261 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     4321 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     4381 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     4441 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     4501 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     4561 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     4621 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     4681 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     4741 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     4801 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     4861 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     4921 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     4981 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     5041 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     5101 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     5161 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     5221 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     5281 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     5341 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     5401 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     5461 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     5521 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     5581 gcgcacattt ccccgaaaag tgccac
//
