LOCUS       sequence-102248-        7726 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  E. coli pACYC based expression vector with N-terminal Hexa-HIS and a
            thrombin protease cleavage site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7726)
  TITLE     Arrowsmith Lab Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7726)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pACYC-LIC+
FEATURES             Location/Qualifiers
     source          1..7726
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(864..1946)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     1908..1927
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1947..2024)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     2230..2249
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2333..2352
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2333..2351
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2352..2376
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             2391..2413
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2433..2450
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2460..2477
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     protein_bind    complement(2496..2529)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2544..2989
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             join(2990..3076,3077..4411)
                     /codon_start=1
                     /gene="Bacillus subtilis sacB"
                     /product="secreted levansucrase that renders bacterial 
                     growth sensitive to sucrose"
                     /label=SacB
                     /note="negative selection marker"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     CDS             4504..4521
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4574..4592)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4588..4635
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(4855..5514)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     5429..5448
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(5515..5617)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(6009..6026)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6143..6687)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(6223..6242)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(6918..6923,6924..6941,6942..6948)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    6956..6972
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     6961..6983
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6980..6996
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6980..6996
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 cgccgctagg tgtgcgagtt caagtctcgc ctcccgcacc attcaccaga aagcgttgat
       61 cggatgccct cgagtcgggc agcgttgggt cctggccacg ggtgcgcatg atcgtgctcc
      121 tgtcgttgag gacccggcta ggctggcggg gttgccttac tggttagcag aatgaatcac
      181 cgatacgcga gcgaacgtga agcgactgct gctgcaaaac gtctgcgacc tgagcaacaa
      241 catgaatggt cttcggtttc cgtgtttcgt aaagtctgga aacgcggaag tcccctacgt
      301 gctgctgaag ttgcccgcaa cagagagtgg aaccaaccgg tgataccacg atactatgac
      361 tgagagtcaa cgccatgagc ggcctcattt cttattctga gttacaacag tccgcaccgc
      421 tgccggtagc tccttccggt gggcgcgggg catgactatc gtcgccgcac ttatgactgt
      481 cttctttatc atgcaactcg taggacaggt gccggcagcg cccaacagtc ccccggccac
      541 ggggcctgcc accataccca cgccgaaaca agcgccctgc accattatgt tccggatctg
      601 catcgcagga tgctgctggc taccctgtgg aacacctaca tctgtattaa cgaagcgcta
      661 accgttttta tcaggctctg ggaggcagaa taaatgatca tatcgtcaat tattacctcc
      721 acggggagag cctgagcaaa ctggcctcag gagtcatgcc ccgcgcccac cggaaggagc
      781 tgactgggtt gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct
      841 aacttacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
      901 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg
      961 gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac cgcctggccc
     1021 tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg
     1081 atggtggtta acggcgggat ataacatgag ctgtcttcgg tatcgtcgta tcccactacc
     1141 gagatatccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc
     1201 atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag catttgcatg
     1261 gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat cggctgaatt
     1321 tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga gacagaactt
     1381 aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg ctccacgccc
     1441 agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg gtcagagaca
     1501 tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca
     1561 tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc
     1621 gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt
     1681 tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg
     1741 gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg
     1801 ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg
     1861 tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg
     1921 acatcgtata acgttactgg tttcacattc accaccctga attgactctc ttccgggcgc
     1981 tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc
     2041 tcccttatgc gactcctgca ttaggaagca gcccagtagt aggttgaggc cgttgagcac
     2101 cgccgccgca aggaatggtg catgcaagga gatggcgccc aacagtcccc cggccacggg
     2161 gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc
     2221 ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat
     2281 gccggccacg atgcgtccgg cgtagaggat cgagatctcg atcccgcgaa attaatacga
     2341 ctcactatag gggaattgtg agcggataac aattcccctc tagaaataat tttgtttaac
     2401 tttaagaagg agatatacat atgggcagca gccatcatca tcatcatcac agcagcggcc
     2461 tggttccgcg tggtagtaga gaccctgaaa gatccataac ttcgtatagc atacattata
     2521 cgaagttatg cggccgcgac gtccacatat acctgccgtt cactattatt tagtgaaatg
     2581 agatattatg atattttctg aattgtgatt aaaaaggcaa ctttatgccc atgcaacaga
     2641 aactataaaa aatacagaga atgaaaagaa acagatagat tttttagttc tttaggcccg
     2701 tagtctgcaa atccttttat gattttctat caaacaaaag aggaaaatag accagttgca
     2761 atccaaacga gagtctaata gaatgaggtc gaaaagtaaa tcgcgcgggt ttgttactga
     2821 taaagcaggc aagacctaaa atgtgtaaag ggcaaagtgt atactttggc gtcacccctt
     2881 acatatttta ggtctttttt tattgtgcgt aactaacttg ccatcttcaa acaggagggc
     2941 tggaagaagc agaccgctaa cacagtacat aaaaaaggag acatgaacga tgaacatcaa
     3001 aaagtttgca aaacaagcaa cagtattaac ctttactacc gcactgctgg caggaggcgc
     3061 aactcaagcg tttgcgaaag aaacgaacca aaagccatat aaggaaacat acggcatttc
     3121 ccatattaca cgccatgata tgctgcaaat ccctgaacag caaaaaaatg aaaaatatca
     3181 agttcctgag ttcgattcgt ccacaattaa aaatatctct tctgcaaaag gcctggacgt
     3241 ttgggacagc tggccattac aaaacgctga cggcactgtc gcaaactatc acggctacca
     3301 catcgtcttt gcattagccg gagatcctaa aaatgcggat gacacatcga tttacatgtt
     3361 ctatcaaaaa gtcggcgaaa cttctattga cagctggaaa aacgctggcc gcgtctttaa
     3421 agacagcgac aaattcgatg caaatgattc tatcctaaaa gaccaaacac aagaatggtc
     3481 aggttcagcc acatttacat ctgacggaaa aatccgttta ttctacactg atttctccgg
     3541 taaacattac ggcaaacaaa cactgacaac tgcacaagtt aacgtatcag catcagacag
     3601 ctctttgaac atcaacggtg tagaggatta taaatcaatc tttgacggtg acggaaaaac
     3661 gtatcaaaat gtacagcagt tcatcgatga aggcaactac agctcaggcg acaaccatac
     3721 gctgagagat cctcactacg tagaagataa aggccacaaa tacttagtat ttgaagcaaa
     3781 cactggaact gaagatggct accaaggcga agaatcttta tttaacaaag catactatgg
     3841 caaaagcaca tcattcttcc gtcaagaaag tcaaaaactt ctgcaaagcg ataaaaaacg
     3901 cacggctgag ttagcaaacg gcgctctcgg tatgattgag ctaaacgatg attacacact
     3961 gaaaaaagtg atgaaaccgc tgattgcatc taacacagta acagatgaaa ttgaacgcgc
     4021 gaacgtcttt aaaatgaacg gcaaatggta cctgttcact gactcccgcg gatcaaaaat
     4081 gacgattgac ggcattacgt ctaacgatat ttacatgctt ggttatgttt ctaattcttt
     4141 aactggccca tacaagccgc tgaacaaaac tggccttgtg ttaaaaatgg atcttgatcc
     4201 taacgatgta acctttactt actcacactt cgctgtacct caagcgaaag gaaacaatgt
     4261 cgtgattaca agctatatga caaacagagg attctacgca gacaaacaat caacgtttgc
     4321 gcctagcttc ctgctgaaca tcaaaggcaa gaaaacatct gttgtcaaag acagcatcct
     4381 tgaacaagga caattaacag ttaacaaata aaaacgcaaa agaaaatgcc gatatcctat
     4441 tggcattgac gtcaggtggc acttttcggt ctcgtgatga cgaagcttgc ggccgcactc
     4501 gagcaccacc accaccacca ctgagatccg gctgctaaca aagcccgaaa ggaagctgag
     4561 ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc
     4621 ttgaggggtt ttttgctgaa acctcaggca tttgagaagc acacggtcac actgcttccg
     4681 gtagtcaata aaccggtaaa ccagcaatag acataagcgg ctatttaacg accctgccct
     4741 gaaccgacga ccgggtcgaa tttgctttcg aatttctgcc attcatccgc ttattatcac
     4801 ttattcaggc gtagcaccag gcgtttaagg gcaccaataa ctgccttaaa aaaattacgc
     4861 cccgccctgc cactcatcgc agtactgttg taattcatta agcattctgc cgacatggaa
     4921 gccatcacag acggcatgat gaacctgaat cgccagcggc atcagcacct tgtcgccttg
     4981 cgtataatat ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt
     5041 taaatcaaaa ctggtgaaac tcacccaggg attggctgag acgaaaaaca tattctcaat
     5101 aaacccttta gggaaatagg ccaggttttc accgtaacac gccacatctt gcgaatatat
     5161 gtgtagaaac tgccggaaat cgtcgtggta ttcactccag agcgatgaaa acgtttcagt
     5221 ttgctcatgg aaaacggtgt aacaagggtg aacactatcc catatcacca gctcaccgtc
     5281 tttcattgcc atacggaatt ccggatgagc attcatcagg cgggcaagaa tgtgaataaa
     5341 ggccggataa aacttgtgct tatttttctt tacggtcttt aaaaaggccg taatatccag
     5401 ctgaacggtc tggttatagg tacattgagc aactgactga aatgcctcaa aatgttcttt
     5461 acgatgccat tgggatatat caacggtggt atatccagtg atttttttct ccattttagc
     5521 ttccttagct cctgaaaatc tcgataactc aaaaaatacg cccggtagtg atcttatttc
     5581 attatggtga aagttggaac ctcttacgtg ccgatcaacg tctcattttc gccaaaagtt
     5641 ggcccagggc ttcccggtat caacagggac accaggattt atttattctg cgaagtgatc
     5701 ttccgtcaca ggtatttatt cggcgcaaag tgcgtcgggt gatgctgcca acttactgat
     5761 ttagtgtatg atggtgtttt tgaggtgctc cagtggcttc tgtttctatc agctgtccct
     5821 cctgttcagc tactgacggg gtggtgcgta acggcaaaag caccgccgga catcagcgct
     5881 agcggagtgt atactggctt actatgttgg cactgatgag ggtgtcagtg aagtgcttca
     5941 tgtggcagga gaaaaaaggc tgcaccggtg cgtcagcaga atatgtgata caggatatat
     6001 tccgcttcct cgctcactga ctcgctacgc tcggtcgttc gactgcggcg agcggaaatg
     6061 gcttacgaac ggggcggaga tttcctggaa gatgccagga agatacttaa cagggaagtg
     6121 agagggccgc ggcaaagccg tttttccata ggctccgccc ccctgacaag catcacgaaa
     6181 tctgacgctc aaatcagtgg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     6241 cccctggcgg ctccctcgtg cgctctcctg ttcctgcctt tcggtttacc ggtgtcattc
     6301 cgctgttatg gccggtttgt ctcattccac gcctgacact cagttccggg taggcagttc
     6361 gctccaagct ggactgtatg cacgaacccc ccgttcagtc cgaccgctgc gccttatccg
     6421 gtaactatcg tcttgagtcc aacccggaaa gacatgcaaa agcaccactg gcagcagcca
     6481 ctggtaattg atttagagga gttagtcttg aagtcatgcg ccggttaagg ctaaactgaa
     6541 aggacaagtt ttggtgactg cgctcctcca agccagttac ctcggttcaa agagttggta
     6601 gctcagagaa ccttcgaaaa accgccctgc aaggcggttt tttcgttttc agagcaagag
     6661 attacgcgca gaccaaaacg atctcaagaa gatcatctta ttaatcagat aaaatatttc
     6721 tagatttcag tgcaatttat ctcttcaaat gtagcacctg aagtcagccc catacgatat
     6781 aagttgtaat tctcatgttt gacagcttat catcgataag cttgcatgca tccatcaaaa
     6841 aaatattgac aacataaaaa actttgtgtt atacttgtaa cgctacatgg agattaactc
     6901 aatctagcta gagaggcttt acactttatg cttccggctc gtataatgtg tggaattgtg
     6961 agcggataac aatttcacac aggaaacagc tatgaccatg attacggatt cactggaact
     7021 ctagaccaaa gagaggacac aatgcagggt tctgtgacag agtttctaaa accgcgcctg
     7081 gttgatatcg agcaagtgag ttcgacgcac gccaaggtga cccttgagcc tttagagcgt
     7141 ggctttggcc atactctggg taacgcactg cgccgtattc tgctctcatc gatgccgggt
     7201 tgcgcggtga ccgaggttga gattgatggt gtactacatg agtacagcac caaagaaggc
     7261 gttcaggaag atatcctgga aatcctgctc aacctgaaag ggctggcggt gagagttcag
     7321 ggcaaagatg aagttattct taccttgaat aaatctggca ttggccctgt gactgcagcc
     7381 gatatcaccc acgacggtga tgtcgaaatc gtcaagccgc agcacgtgat ctgccacctg
     7441 accgatgaga acgcgtctat tagcatgcgt atcaaagttc agcgcggtcg tggttatgtg
     7501 ccggcttcta cccgaattca ttcggaagaa gatgagcgcc caatcggccg tctgctggtc
     7561 gacgcatgct acagccctgt ggagcgtatt gcctacaatg ttgaagcagc gcgtgtagaa
     7621 cagcgtaccg acctggacaa gctggtcatc gaaatggaaa ccaacggcac aatcgatcct
     7681 gaagaggcga ttcgtcgtgc ggcaaccatt ctggctgaac aactgg
//
