LOCUS       Exported                5699 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 5699)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Aug 13, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     Insert from pBD25 pCR8_rap-1 cDNA-mGFP 652 to 2061
            Backbone from pczgy1091 punc-17B dest 2990 to 1307
FEATURES             Location/Qualifiers
     source          1..5699
                     /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
     gene            1959..2675
                     /locus_tag="mGFP"
                     /label=mGFP
     primer_bind     complement(2022..2041)
                     /locus_tag="oSC186 reverse"
                     /label=oSC186 reverse
     misc_feature    2679..2701
                     /locus_tag="3' homology arm"
                     /label=3' homology arm
     primer_bind     complement(join(2679..2684,2693..2694))
                     /locus_tag="Reverse primer"
                     /label=Reverse primer
     misc_feature    2695
                     /locus_tag="PCR insertion between TA(1)"
                     /label=PCR insertion between TA
                     /label=PCR insertion between TA(1)
     misc_recomb     complement(2711..2717)
                     /locus_tag="attB2"
                     /label=attB2
     misc_feature    2718..2732
                     /locus_tag="attR2"
                     /label=attR2
     misc_recomb     complement(2718..2731)
                     /locus_tag="attB2(1)"
                     /label=attB2
                     /label=attB2(1)
     misc_feature    complement(2718..2724)
                     /locus_tag="Gateway Recomb2"
                     /label=Gateway Recomb2
     3'UTR           2809..3492
                     /locus_tag="unc-54 3'UTR"
                     /label=unc-54 3'UTR
     primer_bind     complement(3509..3528)
                     /locus_tag="T3"
                     /label=T3
     primer_bind     complement(3546..3566)
                     /locus_tag="M13-rev"
                     /label=M13-rev
     misc_binding    complement(3572..3594)
                     /locus_tag="LacO"
                     /label=LacO
     rep_origin      complement(3934..4616)
                     /direction=LEFT
                     /locus_tag="ColE1 origin"
                     /label=ColE1 origin
     CDS             complement(4714..5373)
                     /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 ggtgagcaag ggcgaggagc
     1981 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     2041 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
     2101 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
     2161 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     2221 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
     2281 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     2341 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     2401 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga
     2461 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     2521 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccaagc
     2581 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     2641 ccgggatcac tctcggcatg gacgagctgt acaagtagca tatggcggcc gcggaagggc
     2701 gaattcgacc cagctttctt gtacaaagtg gtgatatcga attcctgcag cccgggggat
     2761 ccactagttc tagagcggcc gccaccgcgg ccgctgtcat cagatcgcca tctcgcgccc
     2821 gtgcctctga cttctaagtc caattactct tcaacatccc tacatgctct ttctccctgt
     2881 gctcccaccc cctatttttg ttattatcaa aaaacttctc ttaatttctt tgttttttag
     2941 cttcttttaa gtcacctcta acaatgaaat tgtgtagatt caaaaataga attaattcgt
     3001 aataaaaagt cgaaaaaaat tgtgctccct ccccccatta ataataattc tatcccaaaa
     3061 tctacacaat gttctgtgta cacttcttat gttttttact tctgataaat ttttttgaaa
     3121 catcatagaa aaaaccgcac acaaaatacc ttatcatatg ttacgtttca gtttatgacc
     3181 gcaattttta tttcttcgca cgtctgggcc tctcatgacg tcaaatcatg ctcatcgtga
     3241 aaaagttttg gagtattttt ggaatttttc aatcaagtga aagtttatga aattaatttt
     3301 cctgcttttg ctttttgggg tttcccctat tgtttgtcaa gatttcgagg acggcgtttt
     3361 tcttgctaaa atcacaagta ttgatgagca cgatgcaaga aagatcggaa gaaggtttgg
     3421 gtttgaggct cagtggaagg tgagtagaag ttgataattt gaaagtggag tagtgtctat
     3481 ggggtttttg ccgagctcca gcttttgttc cctttagtga gggttaattg cgcgcttggc
     3541 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     3601 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     3661 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     3721 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     3781 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     3841 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     3901 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     3961 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     4021 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4081 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4141 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     4201 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     4261 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     4321 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     4381 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     4441 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     4501 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     4561 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     4621 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     4681 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     4741 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     4801 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     4861 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     4921 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     4981 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     5041 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     5101 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     5161 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     5221 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     5281 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     5341 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     5401 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     5461 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     5521 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     5581 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     5641 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccac
//
