LOCUS       p5E_(-3kb)nrg1_t        5754 bp DNA     circular SYN 03-MAR-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5754)
  AUTHORS   dacer
  TITLE     Direct Submission
  JOURNAL   Exported Mar 3, 2023 from SnapGene Viewer 6.2.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5754
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      268..295
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      387..473
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    593..688
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    691..711
                     /label=MCS
                     /note="/ApEinfo_fwdcolor=cyan
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
                     /ApEinfo_revcolor=green"
     primer_bind     complement(691..711)
                     /label=P4_P1R_R1
     primer_bind     695..731
                     /label=T2_3055_G_F1
     primer_bind     695..712
                     /label=T2_N_Gbloc_G_F
     misc_feature    712..3775
                     /label=T2_3055_cons
     misc_feature    712..731
                     /label=3055
     primer_bind     712..731
                     /label=T2_3055_F
     primer_bind     934..957
                     /label=T2_N_Gbloc_G_F
     primer_bind     1134..1149
                     /label=T2_2014_F
     primer_bind     1185..1199
                     /label=T2_2524_F
     primer_bind     1645..1665
                     /label=T2_2067_F
     misc_feature    1698..1716
                     /label=2014
     primer_bind     1698..1716
                     /label=T2_2014_F
     misc_feature    1729..1749
                     /label=1983
     primer_bind     1729..1749
                     /label=T2_1983_F
     primer_bind     2000..2021
                     /label=T2_1707_F
     primer_bind     complement(2328..2348)
                     /label=1677_R
     primer_bind     2328..2348
                     /label=1399_F
     misc_feature    2563..2587
                     /label=minus_F
     primer_bind     2563..2587
                     /label=T2_7kb_F6_60C
     misc_feature    2563..2584
                     /label=minus_F_57
     primer_bind     2563..2584
                     /label=T2_7_F6
     primer_bind     2563..2584
                     /label=T2_7kb_F5
     primer_bind     3021..3036
                     /label=T2_700bp_F1
     primer_bind     3394..3416
                     /label=T2p_F2
     primer_bind     3398..3419
                     /label=T2p_F1
     primer_bind     3531..3550
                     /label=T2p_F3
     primer_bind     3590..3605
                     /label=T2_1kb_R7_to_ATG_SNP_C
     primer_bind     3614..3631
                     /label=TII-R
     primer_bind     3665..3685
                     /label=TII-R2
     primer_bind     complement(3708..3731)
                     /label=T2_1kb_R6
     primer_bind     complement(3711..3739)
                     /label=T2_1kb_R5
     primer_bind     complement(3761..3812)
                     /label=T2_1kb_R7_to_ATG_SNP_A
     primer_bind     complement(3761..3812)
                     /label=T2_1kb_R7_to_ATG_SNP_C
     primer_bind     complement(3761..3788)
                     /label=T2_R9_xho
     primer_bind     complement(3761..3775)
                     /label=T2_1_R8
     primer_bind     complement(3764..3792)
                     /label=T2_G_R8
     primer_bind     complement(3764..3788)
                     /label=T2_R8_xho
     misc_feature    3776..3798
                     /label=MCS
                     /note="/ApEinfo_fwdcolor=cyan
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
                     /ApEinfo_revcolor=green"
     primer_bind     3776..3794
                     /label=P4_P1R_F1
     primer_bind     3777..3791
                     /label=T2_7kb_F5
     primer_bind     3777..3791
                     /label=T2_7kb_F6_60C
     primer_bind     complement(3797..3812)
                     /label=T2_1kb_R6
     primer_bind     complement(3798..3812)
                     /label=T2_1kb_R5
     protein_bind    3799..3923
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    3800..3956
                     /label=attR1
                     /note="/ApEinfo_fwdcolor=#ffff80
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
                     /ApEinfo_revcolor=#ff0080"
     promoter        complement(3996..4014)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4019..4035)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             4148..4957
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      5104..5692
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccgc gttaacgcta ccatggagct ccaaataatg
      601 attttatttt gactgatagt gacctgttcg ttgcaacaaa ttgataagca atgctttttt
      661 ataatgccaa ctttgtatag aaaagttgct ggatcccatc gattcgaatt cccactgagc
      721 caccatgtag cctatgtttt atgttcaatc cacttaaatt tgtaaatatg tgtaagttaa
      781 ctgatttgtg ttgggacaac aggaaggaat tatgtggaac ccaacatgtt ttaacagttg
      841 ttcaatttac atgtagaaat tagttccaag accaatgctt gaacacaact aggttttaga
      901 aactgaccca attaaaaaag gacaactttg tttcaattgt gcttgtgtta aagctgcctt
      961 acaaagatgt acagggtaat aattaaagag aaagttcttt attgtggaat atcttcaata
     1021 taaacggtat caagccaaaa tgtggggggg ccctggtgaa gtcagttgtt agcactgttg
     1081 ctaccaagcc agtcagtttc tgcgccaagt ttacacgttc tctctgtgct caagtgggtt
     1141 tcccccaggt tccccagttt cctcctacca tccaaaaaca tacaccagag acatgctcct
     1201 agtaagtagt tatctcttaa gagcattcac tatctgttca ttatctacta cagcagggga
     1261 gttctcgaga tttacctgag cccaaactct cctttcgctt tgcaaatggg agagagcccc
     1321 ggactcgagg atcttatgag ctcagggctc tctcccggga ccgcatgcca aacaagcttg
     1381 aagtaagaat gattctaagg gcctacgccg ttttagggcc cccagagata ggatgagggg
     1441 cccatcacaa cccccccccc tcttcatttt tatccgcaac acccatgaaa tgttttctta
     1501 gggcctgtac cggccagcgg cgcccctgtc tggataaaaa atgcttttct gacttgattt
     1561 tttatttttt ttgtcttgtg tttgtctagt tcaaaaatgg tagcatggtg actcattggt
     1621 tcgcactgtt gcctaacagc aagaagatcg ctggtttaag tcccggctgg tccgtttttg
     1681 tgtggagatt gcatggtcgt gtgggtttcc tccaggtgct tcgatttccc ccacattcca
     1741 cagtcatgca ctataggtgg attgcatgaa ctaaactagc tgtagtgtat gagagtgtgt
     1801 gtgaatgata gtgtgtatac gtgtttccta gtactgggtt gctgtggcaa tcccagatta
     1861 acaaaaggat gaataattag ccaataattt attagcttga gggacatgat agcattgaaa
     1921 ctgaacatat atcgactgat aactaattta gccaccaata tattgtttat aaccgccaaa
     1981 aggtttaaca gaagtgcctg gattgcagac atggcaaacg ctttggacgt tatctcccag
     2041 aacactgatt tgctgtaact gcagcctaaa ttaagacatt tccagcacct ctttaaaata
     2101 actgtccttt tccagagcac gtcctgaatt cccaagtaat ggccctttta gaatcagttg
     2161 gtgttagcaa ctctttcact ccatgcttta aataatacac agtctaaatt ggcacggaca
     2221 taaaagtcac attatgaaac aaaaagcagc ttaattttgc agattattat ttggtttcac
     2281 taggttaata caccacacaa gagttctcaa ctccggcccc agagaccccc tttcatgcag
     2341 agttcagctc cataatcaaa cacacctgaa ccaactaatc catgtgtcaa tacacacact
     2401 agcagtctta ggagttttct cactaaattg acactttcac taatatactg ctattatttg
     2461 agtctaaatg tatccctgta tttattttca ggacctcaca tgtgctaaac ttgggtaaca
     2521 ctttttattt tgatggtcca cttgagtatt agtagactgt ctgcttagca actgttgata
     2581 ctgctccttc aacagacatt taactgacta taagaaactt tgcaagtaca gttgaagtca
     2641 gaatatcccc cccccccctg tttattttcc ccccaatttc tgtgaacgga cagctgattt
     2701 ttttcaacac atttttaaat gtgaaaaaat attattagaa atactttgaa aatttctgca
     2761 ttgatcctga gggccagcct gaacacccac tagtgaccta cttacacctg ctgcttctcc
     2821 acgatagacg tccaatgttc tccagcctcc tgtgcctaga ctgcaggtct gcacaagatg
     2881 tttggccaga ggagaaattg tcgtgcccaa ctgagtctgg tttctctcaa ggtcttttcc
     2941 ttcacttttg tcaattggtg aagttatgtt tctcgccact gtccccactg gcttgcttcc
     3001 tgttgaaaaa atccagctta aaccagccta agctggttta agctggattt ttcagcaggg
     3061 ttggtttggg acttgtggag ctgcgcaatg atggatttgt tcttcattgt ttggactttc
     3121 agcagtgaaa aattaaacca cactgaactg aattaaactg aacttcaact ctaaaaactg
     3181 gactgacggt ttcaattcac tagaacttct atgttaggct gctttgaccc aatcattgta
     3241 aaagcgttaa agaaaaaaaa attaaattga aagctaatat gttcttgata aaatgtccct
     3301 aaatagacat aaacatgttt agagacagtt tatttaattt ttattattat ttaaatttaa
     3361 gtaatattat ttgaaaaatg atcactttta tatcagaggc aatcatctat atgccgacga
     3421 agattattgt tgactatttc atatttaaac agtggttttc ttttttcata ttagcctact
     3481 gaagtacaca gtacttactc atatttagta gctatgttca cattgaaaat cccgtcccac
     3541 ttaattgacc ataagctggc aataattagc gtgattccac agtggaccgc caccccgcgc
     3601 tgtccttggt cgcggtgtcg gtggggaaat cctgctcacc ctcggctggg atctttctca
     3661 ccccgcctcc tgtaatacga tcagctgaag ggctaacaga ccccacgcgc ctcttcattc
     3721 actttttctg ctccacttta attcatccag ctaaacgcaa ggaccgagcg cggaggagcc
     3781 tctagaaccg gtggtaccgc aagtttgtac aaaaaagttg aacgagaaac gtaaaatgat
     3841 ataaatatca atatattaaa ttagattttg cataaaaaac agactacata atactgtaaa
     3901 acacaacata tgcagtcact atgaaccaac tacttagatg gtattagtga cctgtagaat
     3961 tcgagctcta gagctgcagg gcggccgcga tatcccctat agtgagtcgt attacatggt
     4021 catagctgtt tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca
     4081 agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag
     4141 gggtgttatg agccatattc aacgggaaac gtcgaggccg cgattaaatt ccaacatgga
     4201 tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat
     4261 ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag
     4321 cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc
     4381 tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc
     4441 gatccccgga aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat
     4501 tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc
     4561 ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt
     4621 ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa
     4681 agaaatgcat aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc
     4741 acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt
     4801 cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc
     4861 tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa
     4921 attgcagttt catttgatgc tcgatgagtt tttctaatca gaattggtta attggttgta
     4981 acactggcag agcattacgc tgacttgacg ggacggcgca agctcatgac caaaatccct
     5041 taacgtgagt tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     5101 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     5161 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     5221 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     5281 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     5341 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     5401 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     5461 gacctacacc gaactgagat acctacagcg tgagcattga gaaagcgcca cgcttcccga
     5521 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     5581 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     5641 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     5701 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtt
//
