LOCUS       Exported                6998 bp ds-DNA     circular SYN 06-SEP-2024
DEFINITION  Gibson/Homology Cloning ligation product.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6998)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Apr 11, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6998
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    1..4115
                     /label=EML002 backbone (fEL077+fEL076) extraction (con...
     misc_feature    1..4115
                     /label=Extracted region from EML002
     CDS             complement(171..1031)
                     /label=AmpR
     promoter        complement(1032..1136)
                     /label=AmpR promoter
     primer_bind     1176..1195
                     /label=oEL180
     mobile_element  complement(1210..1375)
                     /label=transposon:Tn7
     misc_feature    1475..4076
                     /label=oEL172 + oEL159 (concatenated sequence 2)
     misc_feature    1475..1504
                     /label=Ligation
     primer_bind     complement(1475..1504)
                     /label=oEL173
     primer_bind     1475..1504
                     /label=oEL172
     primer_bind     complement(1475..1504)
                     /label=oEL173
     primer_bind     1475..1504
                     /label=oEL172
     primer_bind     complement(1501..1523)
                     /label=oEL236
     CDS             1507..2322
                     /label=KanR
     primer_bind     complement(1717..1736)
                     /label=oEL179
     terminator      2701..2732
                     /label=T4-32
     primer_bind     3660..3679
                     /label=oEL178
     terminator      3756..3787
                     /label=T4-32
     promoter        complement(3891..3968)
                     /label=AmpR promoter
     primer_bind     3994..4022
                     /label=oEL245
     misc_feature    4044..4076
                     /label=Ligation
     primer_bind     complement(4049..4068)
                     /label=oEL182
     primer_bind     complement(4055..4076)
                     /label=3G_bb_R
     primer_bind     complement(4056..4076)
                     /label=oEL159
     misc_feature    4077..5026
                     /label=fOW025 (concatenated sequence 2)
     misc_feature    4077..4120
                     /label=UNS1_A (concatenated sequence 1)
     primer_bind     4077..4116
                     /label=UNS1
     misc_feature    4077..4115
                     /label=Ligation
     misc_feature    4077..4115
                     /label=UNS1
     primer_bind     4077..4115
                     /label=UNS1_F
     primer_bind     complement(4077..4115)
                     /label=UNS1_R
     misc_feature    4116
                     /label=Extracted region from fOW025
     misc_feature    4117..4986
                     /label=P2m-U17m-TurboRFP-t0 (concatenated sequence 2)
     misc_feature    4117..4159
                     /label=P2m (MAYBE) (concatenated sequence 1)
     misc_feature    4117..4159
                     /label=Editing History Insertion
     misc_feature    4117..4155
                     /label=Extracted region from P2m (MAYBE)
     misc_feature    4117..4120
                     /label=Ligation
     primer_bind     4117..4120
                     /label=Binds to A sticky end
     promoter        4121..4155
                     /label=J23119
     misc_feature    4156..4190
                     /label=Editing History Insertion
     misc_feature    4156..4190
                     /label=U17m (concatenated sequence 2)
     misc_feature    4156..4186
                     /label=Extracted region from U17m
     misc_feature    4156..4159
                     /label=Ligation
     5'UTR           4160..4186
                     /label=Bujard RBS
     misc_feature    4187..4887
                     /label=TurboRFP (CDS) - removed BsaI cut-site (concate...
     misc_feature    4187..4887
                     /label=TurboRFP (CDS) (concatenated sequence 2)
     misc_feature    4187..4883
                     /label=Extracted region from TurboRFP (CDS) - removed ...
     misc_feature    4187..4190
                     /label=Ligation
     misc_feature    4187
                     /label=Ligation
     misc_feature    4188..4883
                     /locus_tag="TurboRFP sequence"
                     /label=TurboRFP sequence
     primer_bind     4188..4208
                     /label=TurboRFP (CDS) (1 F) - DVLA backbone w CDS over...
     primer_bind     complement(4786..4814)
                     /label=remove_BsaI_pt2_R
     primer_bind     4811..4834
                     /label=remove_BsaI_pt1_F
     misc_feature    4811
                     /label=G
     misc_feature    4814
                     /label=C
     primer_bind     complement(4864..4883)
                     /label=TurboRFP (CDS) (696 R) - DVLA backbone w CDS ov...
     misc_feature    join(4884..4887,4187)
                     /label=DVLA backbone w CDS overhangs (concatenated seq...
     misc_feature    4884..4986
                     /label=DVLA backbone w terminator overhangs - EML001 w...
     misc_feature    4884..4986
                     /label=EML001 with UNS - lambda t0 terminator (concate...
     misc_feature    4884..4887
                     /label=Ligation
     misc_feature    4884..4887
                     /label=Ligation
     misc_feature    4884..4887
                     /label=Ligation
     misc_feature    4884..4887
                     /label=Extracted region from T2m
     terminator      complement(4888..4982)
                     /label=lambda t0 terminator
     misc_feature    4888..4982
                     /label=Extracted region from EML001 with UNS
     primer_bind     4888..4909
                     /label=EML001 with UNS - lambda t0 terminator (1 F) - ...
     primer_bind     complement(4959..4982)
                     /label=EML001 with UNS - lambda t0 terminator (95 R) -...
     misc_feature    join(4983..4986,4884..4887)
                     /label=DVLA backbone w terminator overhangs (concatena...
     misc_feature    4983..5026
                     /label=UNSX_E (concatenated sequence 3)
     misc_feature    4983..5026
                     /label=Extracted region from UNSX_E
     misc_feature    4983..4986
                     /label=Ligation
     misc_feature    4983..4986
                     /label=Ligation
     primer_bind     4983..4986
                     /label=Binds to E sticky end
     misc_feature    join(4987..5056,4044..4115)
                     /label=fEL042 (concatenated sequence 3)
     misc_feature    4987..6998
                     /label=EML002 backbone (fEL077+fEL076) extraction (con...
     misc_feature    4987..5026
                     /label=Ligation
     primer_bind     4987..5026
                     /label=UNS10
     misc_feature    4987..5026
                     /label=UNSX
     primer_bind     4987..5026
                     /label=UNSX_F
     misc_feature    5027..8502
                     /label=oEL171 + oEL173 (concatenated sequence 1)
     misc_feature    5027..5056
                     /label=Extracted region from fEL042
     misc_feature    5027..5056
                     /label=Ligation
     primer_bind     5027..5048
                     /label=oEL171
     misc_feature    5057..6998
                     /label=Extracted region from EML001 with UNS
     mobile_element  complement(5107..5305)
                     /label=transposon:Tn7
     primer_bind     complement(5314..5335)
                     /label=oEL188
     primer_bind     5315..5334
                     /label=oEL181
     rep_origin      complement(5387..5775)
                     /direction=LEFT
                     /label=R6K gamma ori
     CDS             5772..5801
                     /label=TcR
     CDS             complement(5946..6317)
                     /label=traJ
     oriT            complement(6350..6459)
                     /direction=LEFT
     rep_origin      complement(6593..6998)
                     /direction=LEFT
                     /label=ori
ORIGIN
        1 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
       61 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
      121 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
      181 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
      241 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
      301 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
      361 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
      421 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
      481 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
      541 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
      601 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
      661 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
      721 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
      781 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
      841 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
      901 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
      961 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     1021 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     1081 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     1141 tttccccgaa aagtgccacc tgaattaatt cgagctcgcc caaacataac aggaagaaaa
     1201 atgccccgct gtgggcggac aaaatagttg ggaactggga ggggtggaaa tggagttttt
     1261 aaggattatt tagggaagag tgacaaaata gatgggaact gggtgtagcg tcgtaagcta
     1321 atacgaaaat taaaaatgac aaaatagttt ggaactagat ttcacttatc tggttggcct
     1381 gcaggggggg ggggggaaag ccacgttgtg tctcaaaatc tctgatgtta cattgcacaa
     1441 gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag taatacaagg
     1501 ggtgttatga gccatattca acgggaaacg tcttgctcga ggccgcgatt aaattccaac
     1561 atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg
     1621 acaatctatc gattgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa
     1681 ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt
     1741 atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc
     1801 actgcgatcc ccgggaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa
     1861 aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat
     1921 tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac
     1981 ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc
     2041 tggaaagaaa tgcataagct tttgccattc tcaccggatt cagtcgtcac tcatggtgat
     2101 ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga
     2161 cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag
     2221 ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg
     2281 aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt ggttaattgg
     2341 ttgtaacact ggcagagcat tacgctgact tgacgggacg gcggctttgt tgaataaatc
     2401 gaacttttgc tgagttgaag gatcagatca cgcatcttcc cgacaacgca gaccgttccg
     2461 tggcaaagca aaagttcaaa atcaccaact ggtccaccta caacaaagct ctcatcaacc
     2521 gtggctccct cactttctgg ctggatgatg gggcgattca ggcctggtat gagtcagcaa
     2581 caccttcttc acgaggcaga cctcagcgcc cccccccccc tgcaggtcga ctagagtcgg
     2641 ggatccggtg attgattgag caagctttat gcttgtaaac cgttttgtga aaaaattttt
     2701 aaaataaaaa aggggacctc tagggtcccc aattaattag taatataatc tattaaaggt
     2761 cattcaaaag gtcatccacc ggatcaattc ccctgctcgc gcaggctggg tgccaagctc
     2821 tcgggtaaca tcaaggcccg atccttggag cccttgccct cccgcacgat gatcgtgccg
     2881 tgatcgaaat ccagatcctt gacccgcagt tgcaaaccct cactgatccg catgcccgtt
     2941 ccatacagaa gctgggcgaa caaacgatgc tcgccttcca gaaaaccgag gatgcgaacc
     3001 acttcatccg gggtcagcac caccggcaag cgccgcgacg gccgaggtct tccgatctcc
     3061 tgaagccagg gcagatccgt gcacagcacc ttgccgtaga agaacagcaa ggccgccaat
     3121 gcctgacgat gcgtggagac cgaaaccttg cgctcgttcg ccagccagga cagaaatgcc
     3181 tcgacttcgc tgctgcccaa ggttgccggg tgacgcacac cgtggaaacg gatgaaggca
     3241 cgaacccagt ggacataagc ctgttcggtt cgtaagctgt aatgcaagta gcgtatgcgc
     3301 tcacgcaact ggtccagaac cttgaccgaa cgcagcggtg gtaacggcgc agtggcggtt
     3361 ttcatggctt gttatgactg tttttttggg gtacagtcta tgcctcgggc atccaagcag
     3421 caagcgcgtt acgccgtggg tcgatgtttg tgtctaacaa ttcgttcaag ccgacgccgc
     3481 ttcgcggcgc ggcttaactc aagcgttaga tgcactaagc acataattgc tcacagccaa
     3541 actatcaggt caagtctgct tttattattt ttaagcgtgc ataataagcc ctacacaaat
     3601 tgggagatat atcatgaaag gctggctttt tcttgttatc gcaatagttg gcgaagtaat
     3661 cgcaacatcc gcattaaaat ctagcgaggg ctttactaag ctgatccggt ggatgacctt
     3721 ttgaatgacc tttaatagat tatattacta attaattggg gaccctagag gtcccctttt
     3781 ttattttaaa aattttttca caaaacggtt tacaagcata aagcttgctc aatcaatcac
     3841 cggatcccga cgggcccggt accgcggccg cgcgaattcg agctcggtac catttatcag
     3901 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     3961 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     4021 acattaacct ataaaaatag gcgtatcacg aggccctttc gtcttcacct cgagaacatt
     4081 actcgcatcc attctcaggc tgtctcgtct cgtctcggag ttgacggcta gctcagtcct
     4141 aggtacagtg ctagctactg aattcattaa agaggagaaa ggtaccaatg agcgagctga
     4201 tcaaggagaa catgcacatg aagctgtaca tggagggcac cgtgaacaac caccacttca
     4261 agtgcacatc cgagggcgaa ggcaagccct acgagggcac ccagaccatg aagatcaagg
     4321 tggtcgaggg cggccctctc cccttcgcct tcgacatcct ggctaccagc ttcatgtacg
     4381 gcagcaaagc cttcatcaac cacacccagg gcatccccga cttctttaag cagtccttcc
     4441 ctgagggctt cacatgggag agaatcacca catacgaaga cgggggcgtg ctgaccgcta
     4501 cccaggacac cagcttccag aacggctgca tcatctacaa cgtcaagatc aacggggtga
     4561 acttcccatc caacggccct gtgatgcaga agaaaacacg cggctgggag gccaacaccg
     4621 agatgctgta ccccgctgac ggcggcctga gaggccacag ccagatggcc ctgaagctcg
     4681 tgggcggggg ctacctgcac tgctccttca agaccacata cagatccaag aaacccgcta
     4741 agaacctcaa gatgcccggc ttccacttcg tggaccacag actggaaaga atcaaggagg
     4801 ccgacaaaga aacgtacgtc gagcagcacg agatggctgt ggccaagtac tgcgacctcc
     4861 ctagcaaact ggggcacaga tgaaggtgac tcctgttgat agatccagta atgacctcag
     4921 aactccatct ggatttgttc agaacgctcg gttgccgccg ggcgtttttt attggtgaga
     4981 atgcttccag gatacataga ttaccacaac tccgagccct tccaccccaa gctagcttgg
     5041 aggacaaatc cgccgctagg agcttgcggc cgccttaaga cgcgtggatc cccgggaatt
     5101 tcgaccccac gcccctcttt aatacgacgg gcaatttgca cttcagaaaa tgaagagttt
     5161 gctttagcca taacaaaagt ccagtatgct ttttcacagc ataactggac tgatttcagt
     5221 ttacaactat tctgtctagt ttaagacttt attgtcatag tttagatcta ttttgttcag
     5281 tttaagactt tattgtccgc ccacacccgc ttacgcaggg catccattta ttactcaacc
     5341 gtaaccgatt ttgccaggtt acgcggctgg tcctctagaa ttcccatgtc agccgttaag
     5401 tgttcctgtg tcactcaaaa ttgctttgag aggctctaag ggcttctcag tgcgttacat
     5461 ccctggcttg ttgtccacaa ccgttaaacc ttaaaagctt taaaagcctt atatattctt
     5521 ttttttctta taaaacttaa aaccttagag gctatttaag ttgctgattt atattaattt
     5581 tattgttcaa acatgagagc ttagtacgtg aaacatgaga gcttagtacg ttagccatga
     5641 gagcttagta cgttagccat gagggtttag ttcgttaaac atgagagctt agtacgttaa
     5701 acatgagagc ttagtacgtg aaacatgaga gcttagtacg tactatcaac aggttgaact
     5761 gctgatcttc agatcctcta cgccggacgc atcgtggccg gatcgggggg ggggggcgct
     5821 cggtcttgcc ttgctcgtcg gtgatgtact tcaccagctc cgcgaagtcg ctcttcttga
     5881 tggagcgcat ggggacgtgc ttggcaatca cgcgcacccc ccggccgttt tagcggctaa
     5941 aaaagtcatg gctctgccct cgggcggacc acgcccatca tgaccttgcc aagctcgtcc
     6001 tgcttctctt cgatcttcgc cagcagggcg aggatcgtgg catcaccgaa ccgcgccgtg
     6061 cgcgggtcgt cggtgagcca gagtttcagc aggccgccca ggcggcccag gtcgccattg
     6121 atgcgggcca gctcgcggac gtgctcatag tccacgacgc ccgtgatttt gtagccctgg
     6181 ccgacggcca gcaggtaggc cgacaggctc atgccggccg ccgccgcctt ttcctcaatc
     6241 gctcttcgtt cgtctggaag gcagtacacc ttgataggtg ggctgccctt cctggttggc
     6301 ttggtttcat cagccatccg cttgccctca tctgttacgc cggcggtagc cggccagcct
     6361 cgcagagcag gattcccgtt gagcaccgcc aggtgcgaat aagggacagt gaagaaggaa
     6421 cacccgctcg cgggtgggcc tacttcacct atcctgcccg gctgacgccg ttggatacac
     6481 caaggaaagt ctacacgaac cctttggcaa aatcctgtat atcgtgcgaa aaaggatgga
     6541 tataccgaaa aaatcgctat aatgaccccg aagcagggtt atgcagcgga aaagcgcttt
     6601 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     6661 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     6721 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     6781 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     6841 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     6901 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     6961 gcaagcagca gattacgcgc agaaaaaaag gatctcaa
//
