LOCUS       sequence-356877-        4707 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses I-SceI from an arabinose promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4707)
  AUTHORS   Lee KZ, Mechikoff MA, Parasa MK, Rankin TJ, Pandolfi P, Fitzgerald 
            KS, Hillman ET, Solomon KV
  TITLE     Repurposing the Homing Endonuclease I-SceI for Positive Selection 
            and Development of Gene-Editing Technologies.
  JOURNAL   ACS Synth Biol. 2022 Jan 21;11(1):53-60. doi: 
            10.1021/acssynbio.1c00340. Epub 2022 Jan 10.
  PUBMED    35007422
REFERENCE   2  (bases 1 to 4707)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1021/acssynbio.1c00340"; journalName: "ACS Synth Biol"; date: 
            "2022-01-21- 21"; volume: "11"; issue: "1"; pages: "53-60"
COMMENT     Sequence Label: pColE1-ISceI
FEATURES             Location/Qualifiers
     source          1..4707
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..285
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     208..227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             328..345
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1108..1125)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1278..1364
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1456..1483
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1502..1593
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1594..1662,1663..2454)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1812..1831)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2625..3213
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3114..3133
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3367..3384
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3399..3539
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3465..3484)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3625..3647
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3803..4681)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
       61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
      121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
      181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg
      241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg ttttttgggc
      301 taacaggagg aattaaccat ggggagccac catcaccatc accatggcag atctatgaaa
      361 aacatcaaaa aaaaccaggt aatgaacctg ggtccgaact ctaaactgct gaaagaatac
      421 aaatcccagc tgatcgaact gaacatcgaa cagttcgaag caggtatcgg tctgatcctg
      481 ggtgatgctt acatccgttc tcgtgatgaa ggtaaaacct actgtatgca gttcgagtgg
      541 aaaaacaaag catacatgga ccacgtatgt ctgctgtacg atcagtgggt actgtccccg
      601 ccgcacaaaa aagaacgtgt taaccacctg ggtaacctgg taatcacctg gggcgcccag
      661 actttcaaac accaagcttt caacaaactg gctaacctgt tcatcgttaa caacaaaaaa
      721 accatcccga acaacctggt tgaaaactac ctgaccccga tgtctctggc atactggttc
      781 atggatgatg gtggtaaatg ggattacaac aaaaactcta ccaacaaatc gatcgtactg
      841 aacacccagt ctttcacttt cgaagaagta gaatacctgg ttaagggtct gcgtaacaaa
      901 ttccaactga actgttacgt aaaaatcaac aaaaacaaac cgatcatcta catcgattct
      961 atgtcttacc tgatcttcta caacctgatc aaaccgtacc tgatcccgca gatgatgtac
     1021 aaactgccga acactatctc ctccgaaact ttcctgaaat aagaattcga agcttggctg
     1081 ttttggtgga tgagagaaga ttttcagcct gatacagatt aaatcagaac gcagaagcgg
     1141 tctgataaaa cagaatttgc ctggcggcag tagcgcggtg gtcccacctg accccatgcc
     1201 gaactcagaa gtgaaacgcc gtagcgccga tggtagtgtg gggtctcccc atgcgagagt
     1261 agggaactgc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt
     1321 ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ggagcggatt
     1381 tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg acgcccgcca taaactgcca
     1441 ggcatcaaat taagcagaag gccatcctga cggatggcct ttttgcgttt ctacaaactc
     1501 ttttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     1561 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     1621 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     1681 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     1741 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     1801 tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc
     1861 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     1921 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     1981 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     2041 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     2101 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     2161 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     2221 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     2281 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     2341 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     2401 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     2461 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     2521 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     2581 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     2641 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     2701 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     2761 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     2821 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     2881 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     2941 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     3001 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     3061 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     3121 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     3181 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     3241 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     3301 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     3361 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt
     3421 acgcatctgt gcggtatttc acaccgcata tggtgcactc tcagtacaat ctgctctgat
     3481 gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc
     3541 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     3601 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     3661 caccgaaacg cgcgaggcag cagatcaatt cgcgcgcgaa ggcgaagcgg catgcataat
     3721 gtgcctgtca aatggacgaa gcagggattc tgcaaaccct atgctactcc gtcaagccgt
     3781 caattgtctg attcgttacc aattatgaca acttgacggc tacatcattc actttttctt
     3841 cacaaccggc acggaactcg ctcgggctgg ccccggtgca ttttttaaat acccgcgaga
     3901 aatagagttg atcgtcaaaa ccaacattgc gaccgacggt ggcgataggc atccgggtgg
     3961 tgctcaaaag cagcttcgcc tggctgatac gttggtcctc gcgccagctt aagacgctaa
     4021 tccctaactg ctggcggaaa agatgtgaca gacgcgacgg cgacaagcaa acatgctgtg
     4081 cgacgctggc gatatcaaaa ttgctgtctg ccaggtgatc gctgatgtac tgacaagcct
     4141 cgcgtacccg attatccatc ggtggatgga gcgactcgtt aatcgcttcc atgcgccgca
     4201 gtaacaattg ctcaagcaga tttatcgcca gcagctccga atagcgccct tccccttgcc
     4261 cggcgttaat gatttgccca aacaggtcgc tgaaatgcgg ctggtgcgct tcatccgggc
     4321 gaaagaaccc cgtattggca aatattgacg gccagttaag ccattcatgc cagtaggcgc
     4381 gcggacgaaa gtaaacccac tggtgatacc attcgcgagc ctccggatga cgaccgtagt
     4441 gatgaatctc tcctggcggg aacagcaaaa tatcacccgg tcggcaaaca aattctcgtc
     4501 cctgattttt caccaccccc tgaccgcgaa tggtgagatt gagaatataa cctttcattc
     4561 ccagcggtcg gtcgataaaa aaatcgagat aaccgttggc ctcaatcggc gttaaacccg
     4621 ccaccagatg ggcattaaac gagtatcccg gcagcagggg atcattttgc gcttcagcca
     4681 tacttttcat actcccgcca ttcagag
//
