LOCUS       Exported                5690 bp ds-DNA     circular SYN 12-SEP-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5690)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Aug 13, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     Insert from pBD24 pCR8_rap-1 cDNA-mCherry 652 to 2052
            Backbone from pczgy1091 punc-17B dest 2990 to 1307
FEATURES             Location/Qualifiers
     source          1..5690
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      complement(135..441)
                     /direction=LEFT
                     /locus_tag="F1 ori"
                     /label=F1 ori
     CDS             complement(460..528)
                     /locus_tag="LacZ alpha"
                     /label=LacZ alpha
     misc_feature    497..502
                     /locus_tag="pvui"
                     /label=pvui
     primer_bind     599..616
                     /locus_tag="M13-fwd"
                     /label=M13-fwd
     primer_bind     625..645
                     /locus_tag="T7"
                     /label=T7
     misc_feature    join(653..658,1155..1166)
                     /locus_tag="pacr-2"
                     /label=pacr-2
     misc_feature    653..658
                     /locus_tag="kpn1"
                     /label=kpn1
     misc_feature    653..658
                     /locus_tag="KPNI"
                     /label=KPNI
     promoter        659..1154
                     /locus_tag="Punc-17 beta"
                     /label=Punc-17 beta
     misc_feature    1155..1293
                     /locus_tag="synthetic intron"
                     /label=synthetic intron
     misc_feature    1155..1160
                     /locus_tag="HINDIII"
                     /label=HINDIII
     misc_feature    1161..1166
                     /locus_tag="sph1"
                     /label=sph1
     misc_feature    1161..1166
                     /locus_tag="hindIII"
                     /label=hindIII
     misc_feature    1301..1307
                     /locus_tag="attR1"
                     /label=attR1
     misc_recomb     1301..1307
                     /locus_tag="attB1(1)"
                     /label=attB1
                     /label=attB1(1)
     misc_recomb     1308..1321
                     /locus_tag="attB1"
                     /label=attB1
     misc_recomb     1308..1321
                     /locus_tag="AttL1"
                     /label=AttL1
     misc_feature    1308..1314
                     /locus_tag="Gateway recomb1"
                     /label=Gateway recomb1
     misc_feature    1326..1352
                     /locus_tag="5' homology arm"
                     /label=5' homology arm
     misc_feature    1338
                     /locus_tag="PCR insertion between TA"
                     /label=PCR insertion between TA
     primer_bind     join(1339..1340,1347..1352)
                     /locus_tag="Forward primer"
                     /label=Forward primer
     CDS             1359..1922
                     /locus_tag="rap-1"
                     /label=rap-1
     primer_bind     1359..1382
                     /locus_tag="oSC188"
                     /label=oSC188
     primer_bind     complement(1903..1922)
                     /locus_tag="oSC189"
                     /label=oSC189
     misc_feature    1929..1958
                     /locus_tag="SGSGGSGGSG linker"
                     /label=SGSGGSGGSG linker
     misc_feature    1959..2666
                     /locus_tag="mCherry"
                     /label=mCherry
     primer_bind     complement(2028..2047)
                     /locus_tag="oSC187 reverse"
                     /label=oSC187 reverse
     misc_feature    2670..2692
                     /locus_tag="3' homology arm"
                     /label=3' homology arm
     primer_bind     complement(join(2670..2675,2684..2685))
                     /locus_tag="Reverse primer"
                     /label=Reverse primer
     misc_feature    2686
                     /locus_tag="PCR insertion between TA(1)"
                     /label=PCR insertion between TA
                     /label=PCR insertion between TA(1)
     misc_recomb     complement(2702..2708)
                     /locus_tag="attB2"
                     /label=attB2
     misc_feature    2709..2723
                     /locus_tag="attR2"
                     /label=attR2
     misc_recomb     complement(2709..2722)
                     /locus_tag="attB2(1)"
                     /label=attB2
                     /label=attB2(1)
     misc_feature    complement(2709..2715)
                     /locus_tag="Gateway Recomb2"
                     /label=Gateway Recomb2
     3'UTR           2800..3483
                     /locus_tag="unc-54 3'UTR"
                     /label=unc-54 3'UTR
     primer_bind     complement(3500..3519)
                     /locus_tag="T3"
                     /label=T3
     primer_bind     complement(3537..3557)
                     /locus_tag="M13-rev"
                     /label=M13-rev
     misc_binding    complement(3563..3585)
                     /locus_tag="LacO"
                     /label=LacO
     rep_origin      complement(3925..4607)
                     /direction=LEFT
                     /locus_tag="ColE1 origin"
                     /label=ColE1 origin
     CDS             complement(4705..5364)
                     /locus_tag="AmpR"
                     /label=AmpR
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 gcgcgtaata cgactcacta tagggcgaat tgggtaccgc
      661 ttttggtttt cacaattttc tggtttttgg gatttttgcg ggaattttga gaggagaaag
      721 ttgggagaga agatattgga atggtccccg gggagctgta catttataca tctagcgcca
      781 aggcaatctt cgtaaaaaat aacaatagag tcatccctca gaacagaaaa gaaacccgcc
      841 cgggggctca aatacacatg tcatttttct tttttcgcct gatttttagc ctaaattagt
      901 gtttagcaat tttttaaggt aactcggtgg agtcatttga actttttaaa ctaaaatttc
      961 agaaaaaatt cttttttttt gtcaaaaaat aattttcccg ccggaatttt tcaaaaaaat
     1021 aatttttatt ttatttttca agttccttgt ttcctcttta tttttttatt ttgcccccaa
     1081 aagttttaca ctaaaatatt taattttcag gtaattcatc gaaagtcttt ctattttccg
     1141 catctcttgt tcaaaagctt gcatgcctgc aggtcgactc tagaggatcc ccgggattgg
     1201 ccaaaggacc caaaggtatg tttcgaatga tactaacata acatagaaca ttttcaggag
     1261 gacccttgga gggtaccggt agaaaaaaag cttgatatca caagtttgta caaaaaagca
     1321 ggctccgaat tcgcccttgg actagtcata tgggtaccat gcgggagtat aagattgttg
     1381 tgctcggatc tggaggagta ggaaaatccg cactgacggt acaattcgtg cagggaattt
     1441 tcgtagagaa gtatgaccca acaattgagg atagttacag aaagcaagtg gaagtcgacg
     1501 gccagcaatg tatgcttgag atactggata cagcaggaac ggagcaattc acagcgatgc
     1561 gtgatttgta catgaagaat ggacaaggat ttgttctcgt ttattcaatt actgctcaat
     1621 cgacattcaa tgatcttatg gatcttcgtg atcagattct tcgtgtcaaa gatacggacg
     1681 aggttccaat gattttggta ggaaataagt gtgatttgga agacgaacga gttgttggaa
     1741 aagatcaggg acagaatctc gccaggcagt ttggatcagc attccttgaa acctctgcaa
     1801 aagcaaagat taacgtcagc gaggtgttct acgacctcgt tcgtcaaata aaccgacgtt
     1861 atcccgaatc tggtcgacgc cagggacaat caaataagca gtgctgctcg tgtgtcatca
     1921 tgggtaccag tggttccgga ggatctggag gatctggtat ggtctcaaag ggtgaagaag
     1981 ataacatggc aattattaaa gagtttatgc gtttcaaggt gcatatggag ggatctgtca
     2041 atgggcatga gtttgaaatt gaaggtgaag gagaaggccg accatatgag ggaacacaaa
     2101 ccgcaaaact aaaggtaact aaaggcggac cattaccatt cgcctgggac atcctctctc
     2161 cacagttcat gtatggaagt aaagcttatg ttaaacatcc ggcagatata ccagattatt
     2221 tgaaactttc attcccggag ggttttaagt gggaacgcgt aatgaatttt gaagacggag
     2281 gagttgttac agtgacgcaa gactcaagcc tccaagatgg agaatttatt tataaagtca
     2341 aacttcgagg aacgaatttc ccctcggatg gacctgttat gcagaagaag actatgggat
     2401 gggaagcttc aagtgaaaga atgtaccctg aagacggtgc tcttaaggga gagattaaac
     2461 aacgtcttaa attgaaagat ggaggacatt acgatgctga ggtgaagaca acttacaaag
     2521 ccaaaaaacc agttcagctg ccaggagcgt acaatgttaa tattaaactg gatatcacct
     2581 cccacaacga ggattacact atcgttgagc aatatgaaag agctgaaggg cggcactcga
     2641 caggtggcat ggatgaattg tataagtagc atatggcggc cgcggaaggg cgaattcgac
     2701 ccagctttct tgtacaaagt ggtgatatcg aattcctgca gcccggggga tccactagtt
     2761 ctagagcggc cgccaccgcg gccgctgtca tcagatcgcc atctcgcgcc cgtgcctctg
     2821 acttctaagt ccaattactc ttcaacatcc ctacatgctc tttctccctg tgctcccacc
     2881 ccctattttt gttattatca aaaaacttct cttaatttct ttgtttttta gcttctttta
     2941 agtcacctct aacaatgaaa ttgtgtagat tcaaaaatag aattaattcg taataaaaag
     3001 tcgaaaaaaa ttgtgctccc tccccccatt aataataatt ctatcccaaa atctacacaa
     3061 tgttctgtgt acacttctta tgttttttac ttctgataaa tttttttgaa acatcataga
     3121 aaaaaccgca cacaaaatac cttatcatat gttacgtttc agtttatgac cgcaattttt
     3181 atttcttcgc acgtctgggc ctctcatgac gtcaaatcat gctcatcgtg aaaaagtttt
     3241 ggagtatttt tggaattttt caatcaagtg aaagtttatg aaattaattt tcctgctttt
     3301 gctttttggg gtttccccta ttgtttgtca agatttcgag gacggcgttt ttcttgctaa
     3361 aatcacaagt attgatgagc acgatgcaag aaagatcgga agaaggtttg ggtttgaggc
     3421 tcagtggaag gtgagtagaa gttgataatt tgaaagtgga gtagtgtcta tggggttttt
     3481 gccgagctcc agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg
     3541 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     3601 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     3661 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     3721 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     3781 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     3841 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3901 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3961 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     4021 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4081 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4141 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4201 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4261 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4321 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4381 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4441 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4501 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     4561 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     4621 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     4681 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4741 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     4801 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     4861 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     4921 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     4981 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     5041 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     5101 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     5161 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     5221 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     5281 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     5341 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     5401 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     5461 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     5521 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     5581 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     5641 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
//
