LOCUS       sequence-166638-        2808 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E.coli cloning and expression vector. Has cloning sites downstream 
            of Tet promoter. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2808)
  TITLE     Coleman Lab plasmids
REFERENCE   2  (bases 1 to 2808)
  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"
COMMENT     Sequence Label: pUS212
FEATURES             Location/Qualifiers
     source          1..2808
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     404..423
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     651..669
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(673..716)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    719..740
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             complement(766..1392)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     1297..1316
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        1408..1463
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    1414..1432
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    1444..1462
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_feature    1479..1499
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      1500..1557
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(1581..1598)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1752..2340)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1832..1851)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2523..2617)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(2638..2808,1..489))
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
ORIGIN
        1 catattggcc acgtttaaat caaaactggt gaaactcacc cagggattgg ctgagacgaa
       61 aaacatattc tcaataaacc ctttagggaa ataggccagg ttttcaccgt aacacgccac
      121 atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg tggtattcac tccagagcga
      181 tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa gggtgaacac tatcccatat
      241 caccagctca ccgtctttca ttgccatacg aaattccgga tgagcattca tcaggcgggc
      301 aagaatgtga ataaaggccg gataaaactt gtgcttattt ttctttacgg tctttaaaaa
      361 ggccgtaata tccagctgaa cggtctggtt ataggtacat tgagcaactg actgaaatgc
      421 ctcaaaatgt tctttacgat gccattggga tatatcaacg gtggtatatc cagtgatttt
      481 tttctccatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa atacgcccgg
      541 tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgcccga tcaactcgag
      601 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
      661 tcacgaggca gaatttcaga taaaaaaaat ccttagcttt cgctaaggat gatttctgga
      721 attcgcggcc gcttctagag atcacggaaa aaggttatgc tgcttttaag acccactttc
      781 acatttaagt tgtttttcta atccgcatat gatcaattca aggccgaata agaaggctgg
      841 ctctgcacct tggtgatcaa ataattcgat agcttgtcgt aataatggcg gcatactatc
      901 agtagtaggt gtttcccttt cttctttagc gacttgatgc tcttgatctt ccaatacgca
      961 acctaaagta aaatgcccca cagcgctgag tgcatataat gcattctcta gtgaaaaacc
     1021 ttgttggcat aaaaaggcta attgattttc gagagtttca tactgttttt ctgtaggccg
     1081 tgtacctaaa tgtacttttg ctccatcgcg atgacttagt aaagcacatc taaaactttt
     1141 agcgttatta cgtaaaaaat cttgccagct ttccccttct aaagggcaaa agtgagtatg
     1201 gtgcctatct aacatctcaa tggctaaggc gtcgagcaaa gcccgcttat tttttacatg
     1261 ccaatacaat gtaggctgct ctacacctag cttctgggcg agtttacggg ttgttaaacc
     1321 ttcgattccg acctcattaa gcagctctaa tgcgctgtta atcactttac ttttatctaa
     1381 tctggacatc attaattcct aatttttgtt gacactctat cattgataga gttattttac
     1441 cactccctat cagtgataga gaaaagtgaa tttaatagta ctagtagcgg ccgctgcagt
     1501 ccggcaaaaa agggcaaggt gtcaccaccc tgcccttttt ctttaaaacc gaaaagatta
     1561 cttcgcgtta tgcaggcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     1621 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     1681 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     1741 ttgctggcgt ttttccacag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     1801 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     1861 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     1921 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     1981 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     2041 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     2101 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     2161 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     2221 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     2281 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     2341 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     2401 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     2461 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ctcgaggctt
     2521 ggattctcac caataaaaaa cgcccggcgg caaccgagcg ttctgaacaa atccagatgg
     2581 agttctgagg tcattactgg atctatcaac aggagtccaa gcgagctcga tatcaaatta
     2641 cgccccgccc tgccactcat cgcagtactg ttgtaattca ttaagcattc tgccgacatg
     2701 gaagccatca caaacggcat gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc
     2761 ttgcgtataa tatttgccca tggtgaaaac gggggcgaag aagttgtc
//
