LOCUS       Exported                2580 bp ds-DNA     circular SYN 29-OCT-2024
DEFINITION  synthetic circular DNA
ACCESSION   urn.local...17d-auwcesf
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2580)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Oct 31, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     Imported using the Genbank importer. File name: pBjh_4428.gb [pL0-I]
            E1 [GGAG TACT]. ApEinfo:methylated:1.
FEATURES             Location/Qualifiers
     source          1..2580
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    complement(4..9)
                     /label=BBb scar
                     /note="ApEinfo_revcolor: green ApEinfo_graphicformat: 
                     arrow_data {{0 1 2 0 0 -1} {} 0}width 5 offset 0 label: BBb
                     scar(4)"
     protein_bind    complement(475..479)
                     /label=CTCF Bind (Short)
                     /note="created_by: User"
     rep_origin      589
                     /label=ORI
                     /note="created_by: User"
     primer_bind     689..708
                     /label=G00101
                     /note="Mismatches: 0 created_by: primer3 annotation_group: 
                     37241076164624321b12827-095f-4d96-89b8-54bfccb22708 %GC: 
                     50.0 Tm: 58.3 Hairpin_Tm: None Self_Dimer_Tm: None 
                     Sequence: ATTACCGCCTTTGAGTGAGC"
     primer_bind     689..708
                     /label=G00101
                     /note="created_by: User Sequence: ATTACCGCCTTTGAGTGAGC"
     primer_bind     689..708
                     /label=KK_G00101
     misc_feature    complement(767..773)
                     /label=SapI
                     /note="Recognition_pattern: GCTCTTC(1/4) annotation_group: 
                     GCTCTTC note: Geneious type: restriction site"
     protein_bind    830..835
                     /label=BsaI Bind
                     /note="created_by: User"
     misc_feature    830..835
                     /label=BsaI
                     /note="Recognition_pattern: GGTCTC(1/5) annotation_group: 
                     GGTCTC note: Geneious type: restriction site"
     misc_feature    837..840
                     /label=pL0 I
                     /label=nonstandard type:  Gg_overhang
                     /note="created_by: User"
     misc_feature    841..1167
                     /label=E1 Insulator
                     /label=nonstandard type:  Insulator
                     /note="created_by: User"
     primer_bind     841..862
                     /label=RJ_00346Fa
                     /note="Sequence: GCCACCTTTGGTCTTGACATTC Hairpin_Tm: 46.0 
                     Tm: 60.0 Self_Dimer_Tm: 3.1 %GC: 50.0 created_by: primer3 
                     Extension: GCGAACCATGGATTC modified_by: User"
     protein_bind    919..923
                     /label=CTCF Bind (Short)
                     /note="created_by: User"
     protein_bind    complement(931..935)
                     /label=CTCF Bind (Short)
                     /note="created_by: User"
     protein_bind    985..998
                     /label=Class E CTCF Target
                     /note="created_by: User modified_by: User"
     protein_bind    complement(990..994)
                     /label=CTCF Bind (Short)
                     /note="created_by: User"
     misc_feature    1084..1089
                     /label=XbaI
                     /note="Recognition_pattern: T^CTAGA annotation_group: 
                     TCTAGA note: Geneious type: restriction site"
     primer_bind     1110..1129
                     /label=RJ_00377Fb
                     /note="Sequence: CACCATGGCTTCCTCTTTGG Hairpin_Tm: 37.1 Tm: 
                     58.8 Self_Dimer_Tm: None %GC: 55.0 created_by: primer3"
     misc_feature    1112..1117
                     /label=NcoI
                     /note="Recognition_pattern: C^CATGG annotation_group: 
                     CCATGG note: Geneious type: restriction site"
     misc_feature    complement(1148..1167)
                     /label=RJ_00346Ra
                     /note="Sequence: ATCAGCGGTGTCAGTCTTCC Hairpin_Tm: None Tm: 
                     59.8 Self_Dimer_Tm: 15.2 %GC: 55.0 created_by: primer3 
                     Extension: TCGTGGTCTCAAGCG modified_by: User note: Geneious
                     type: primer_bind_reverse"
     misc_feature    1149..1154
                     /label=BbsI
                     /note="Recognition_pattern: GAAGAC(2/6) annotation_group: 
                     GAAGAC note: Geneious type: restriction site"
     misc_feature    1168..1171
                     /label=pL0 I-P
                     /label=nonstandard type:  Gg_overhang
                     /note="created_by: User"
     protein_bind    complement(1173..1178)
                     /label=BsaI Bind
                     /note="created_by: User"
     misc_feature    complement(1173..1178)
                     /label=BsaI
                     /note="Recognition_pattern: GGTCTC(1/5) annotation_group: 
                     GGTCTC note: Geneious type: restriction site"
     misc_feature    1187..1193
                     /label=SapI
                     /note="Recognition_pattern: GCTCTTC(1/4) annotation_group: 
                     GCTCTTC note: Geneious type: restriction site"
     primer_bind     complement(1200..1218)
                     /label=KK_lvl0Rev
     misc_feature    1328..1351
                     /label=Pamp
                     /note="created_by: User note: Geneious type: 
                     Promoter_prokaryotic"
     misc_feature    complement(1343..1363)
                     /label=JH856R
                     /note="created_by: User Sequence: CAGGGTTATTGTCTCATGAGC 
                     note: Geneious type: primer_bind_reverse"
     misc_feature    complement(1343..1363)
                     /label=JH856R
                     /note="Mismatches: 0 created_by: primer3 annotation_group: 
                     372410859156906c8e796de-2ec0-4dc5-9ceb-b9c822ec2370 %GC: 
                     47.6 Tm: 56.4 Hairpin_Tm: None Self_Dimer_Tm: 13.2 
                     Sequence: CAGGGTTATTGTCTCATGAGC note: Geneious type: 
                     primer_bind_reverse"
     CDS             1398..1557
                     /label=Spec
                     /note="created_by: User"
     misc_feature    1486..1487
                     /label=missing TT
     protein_bind    complement(1621..1625)
                     /label=CTCF Bind (Short)
                     /note="created_by: User"
     misc_feature    complement(1871..1894)
                     /label=RJ_00189Ra
                     /note="Mismatches: 0 created_by: primer3 annotation_group: 
                     372410971476436a042c3c9-eaca-4830-8110-45f44da0096b %GC: 
                     45.8 Tm: 60.3 Hairpin_Tm: 36.5 Self_Dimer_Tm: 4.8 Sequence:
                     CACAACAATGGTGACTTCTACAGC note: Geneious type: 
                     primer_bind_reverse"
     primer_bind     1880..1900
                     /label=RJ_00190Fa
                     /note="Mismatches: 0 created_by: primer3 annotation_group: 
                     372411066187318212241b1-26da-42d6-a894-eaaafa2e0b4c %GC: 
                     52.4 Tm: 60.6 Hairpin_Tm: None Self_Dimer_Tm: 27.4 
                     Sequence: GTCACCATTGTTGTGCACGAC"
     misc_feature    2098..2103
                     /label=BBb scar
                     /note="ApEinfo_revcolor: green ApEinfo_graphicformat: 
                     arrow_data {{0 1 2 0 0 -1} {} 0}width 5 offset 0"
     misc_feature    2475..2480
                     /label=BBb scar
                     /note="ApEinfo_revcolor: green ApEinfo_graphicformat: 
                     arrow_data {{0 1 2 0 0 -1} {} 0}width 5 offset 0 label: BBb
                     scar(1)"
     misc_feature    complement(2487..2492)
                     /label=BBb scar
                     /note="ApEinfo_revcolor: green ApEinfo_graphicformat: 
                     arrow_data {{0 1 2 0 0 -1} {} 0}width 5 offset 0 label: BBb
                     scar(3)"
     rep_origin      complement(join(2505..2580,1..607))
                     /direction=LEFT
                     /label=ColE1 origin
                     /note="ApEinfo_fwdcolor: gray50 ApEinfo_revcolor: gray50 
                     ApEinfo_graphicformat: arrow_data {{0 1 2 0 0 -1} {} 
                     0}width 5 offset 0"
     misc_feature    2573..2578
                     /label=BBb scar
                     /note="ApEinfo_revcolor: green ApEinfo_graphicformat: 
                     arrow_data {{0 1 2 0 0 -1} {} 0}width 5 offset 0 label: BBb
                     scar(2)"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tcactctgtg gtctcaggag
      841 gccacctttg gtcttgacat tcactaagga gtgggcctgg aataaaacca gaaatcccca
      901 ttctcagtct gccccaagcc ctccaattcc gagggcgctg tatgtataag ctcgggcagg
      961 caaaagtcga ctgtgagaac acgccagcag agggcgctgt ggccccatca gtccctgccc
     1021 actgaactct ccagaaggaa aagcggcaag gatgcaaaca agaaaatcag accagactgg
     1081 gagtctagac cctgcgtccc acgcggtccc accatggctt cctctttgga attttcaggg
     1141 ggacccagga agactgacac cgctgattac ttgagaccac gaagtggctc ttcagtggac
     1201 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     1261 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     1321 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     1381 attgaaaaag gaagagtatg cgctcacgca actggtccag aaccttgacc gaacgcagcg
     1441 gtggtaacgg cgcagtggcg gttttcatgg cttgttatga ctgttttttt ttggggtaca
     1501 gtctatgcct cgggcatcca agcagcaagc gcgttacgcc gtgggtcgat gtttgatgtt
     1561 atggagcagc aacgatgtta cgcagcaggg cagtcgccct aaaacaaagt taaacatcat
     1621 gagggaagcg gtgatcgccg aagtatcgac tcaactatca gaggtagttg gcgtcatcga
     1681 gcgccatctc gaaccgacgt tgctggccgt acatttgtac ggctccgcag tggatggcgg
     1741 cctgaagcca cacagcgata ttgatttgct ggttacggtg accgtaaggc ttgatgaaac
     1801 aacgcggcga gctttgatca acgacctttt ggaaacttcg gcttcccctg gagagagcga
     1861 gattctccgc gctgtagaag tcaccattgt tgtgcacgac gacatcattc cgtggcgtta
     1921 tccagctaag cgcgaactgc aatttggaga atggcagcgc aatgacattc ttgcaggtat
     1981 cttcgagcca gccacgatcg acattgatct ggctatcttg ctgacaaaag caagagaaca
     2041 tagcgttgcc ttggtaggtc cagcggcgga ggaactcttt gatccggttc ctgaacagga
     2101 tctatttgag gcgctaaatg aaaccttaac gctatggaac tcgccgcccg actgggctgg
     2161 cgatgagcga aatgtagtgc ttacgttgtc ccgcatttgg tacagcgcag taaccggcaa
     2221 aatcgcgccg aaggatgtcg ctgccgactg ggcaatggag cgcctgccgg cccagtatca
     2281 gcccgtcata cttgaagcta gacaggctta tcttggacaa gaagaagatc gcttggcctc
     2341 gcgcgcagat cagttggaag aatttgtcca ttacgtaaaa ggcgagatca ccaaggtagt
     2401 cggcaaataa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     2461 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     2521 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
//
