LOCUS       pJMP3653        5599 bp DNA     circular SYN 30-MAY-2024
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5599)
  AUTHORS   Jason Peters Lab
  TITLE     Direct Submission
  JOURNAL   Exported May 30, 2024 from SnapGene 7.2.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5599
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        27..104
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             105..1187
                     /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"
     misc_feature    join(1344..1350,1351..1366,1367..1372)
                     /label=PabstBR promoter
     misc_feature    1351..1366
                     /label=lac operator
     misc_feature    1374..1403
                     /label=Gibson ends for NcoI/BamHI cloning
     misc_feature    1392..1403
                     /label=strong RBS
     misc_feature    1407..1463
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     misc_feature    1429..1458
                     /label=Gibson ends for NcoI/BamHI cloning
     terminator      1546..1871
                     /label=rrnB terminators
                     /label=rrnB
     terminator      1666..1752
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1844..1871
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        join(1947..1952,1953..1969,1970..1975)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             2086..2880
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     misc_feature    2977..4318
                     /label=A. baumannii pWH1266 origin
                     /note="from pSGA-km"
     misc_feature    4377..4612
                     /label=expanded oriT
     oriT            4503..4612
                     /note="incP origin of transfer"
     rep_origin      4995..5540
                     /direction=RIGHT
                     /label=E. coli p15A origin
                     /note="medium-copy-number p15A origin of replication can be
                     propagated in E. coli"
ORIGIN
        1 aaggcgaagc ggcatgcatt tacgttgaca ccatcgaatg gtgcaaaacc tttcgcggta
       61 tggcatgata gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt
      121 tatacgatgt cgcagagtat gccggtgtct cttatcagac cgtttcccgc gtggtgaacc
      181 aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg gcggagctga
      241 attacattcc caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg ctgattggcg
      301 ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc
      361 gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag
      421 cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg atcattaact
      481 atccgctgga tgaccaggat gccattgctg tggaagctgc ctgcactaat gttccggcgt
      541 tatttcttga tgtctctgac cagacaccca tcaacagtat tattttctcc catgaagacg
      601 gtacgcgact gggcgtggag catctggtcg cattgggtca ccagcaaatc gcgctgttag
      661 cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat aaatatctca
      721 ctcgcaatca aattcagccg atagcggaac gggaaggcga ctggagtgcc atgtccggtt
      781 ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca
      841 acgatcagat ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg
      901 cggatatctc ggtagtggga tacgacgata ccgaagacag ctcatgttat atcccgccgt
      961 taaccaccat caaacaggat tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc
     1021 aactctctca gggccaggcg gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa
     1081 gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     1141 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     1201 aatgtgagtt agcgcgaatt gatctggttt gacagcttat catcgactgc acggtgcacc
     1261 aatgcttctg gcgtcaggca gccatcggaa gctgtggtat ggctgtgcag gtcgtaaaga
     1321 cgtctcatta atcagataaa atattgacaa tgtgagcgga taacaagata ctgagcacat
     1381 cagcagcaca caggaaacag accatggaat tcgagctcgg tacccgggga tcctctagag
     1441 tcgacctgca ggcatgcaag cttggctgtt ttggcggatg agagaagatt ttcagcctga
     1501 tacagattaa atcagaacgc agaagcggtc tgataaaaca gaatttgcct ggcggcagta
     1561 gcgcggtggt cccacctgac cccatgccga actcagaagt gaaacgccgt agcgccgatg
     1621 gtagtgtggg gtctccccat gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag
     1681 gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg
     1741 agtaggacaa atccgccggg agcggatttg aacgttgcga agcaacggcc cggagggtgg
     1801 cgggcaggac gcccgccata aactgccagg catcaaatta agcagaaggc catcctgacg
     1861 gatggccttt ttcctgaacg gccataagaa cgaaggctgt ctgttgaact ctcgagccgt
     1921 ggaaacggat gaaggcacga acccagttga cataagcctg ttcggttcgt aaactgtaat
     1981 gcaagtagcg tatgcgctca aaactggatg gctttcttgc cgccaaggat ctgatggcgc
     2041 aggggatcaa gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     2101 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     2161 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     2221 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     2281 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     2341 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     2401 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     2461 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     2521 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     2581 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     2641 gtgacccacg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     2701 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     2761 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     2821 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     2881 gcgggactct ggggttcgaa atgactcgag catcagcttc aaaagcgctc cgatgtactg
     2941 cggtttctgg actagtccgt tgacattatc gcagcaggat tttaacattt tgcgttgttc
     3001 caaaagttat caacagccta gaacgtcata ggaagcgatt acagacactt tatagctatc
     3061 agcatgggaa cataagggca ggatgaaata tgggtcttaa aacgcaaatg gtgaggtttt
     3121 agaggtattt tttgaagatg attaaggcgg tttgttttta aaatttttgg cggctctcag
     3181 gctgcttaca ttttaaccag ttcagtgaaa agttcttttt cagcaaattt ctgtttagca
     3241 ccatagctaa aacttgcgtg gaacatatta agaattgacc gaaatgacac aatctcaatt
     3301 atattttttt tgaaaagttt tctttatcaa atattttaaa tcattgattt atatataagt
     3361 atgcattcat tttaataatt aatctttatt taacaatgat ttatctatat tcaattgttt
     3421 aattattctt actaatatta tctctatatc aatatttttt atttaaaaac atatgtttag
     3481 tagtgctttt gattaaagta ccagagggag ggagcagagc tgaatgggaa atactcaccc
     3541 tagagcgatt cttaaaaatc accctaaagt attcccattc gatgtaccgt cggtcggtcg
     3601 ctttcgcatc agggatgaca tcactgtatc aagctgccac tgttatgatt acgattgata
     3661 gcaccgcctg aacacgctca taaccgccaa ttaaatgact actcattgcg tccgctactc
     3721 tgttcagttc ccctagtaat agcgtttttc cgatgtgtgc tagcgtcact gtacctcatc
     3781 acccacacat ggacagattg agttacagac attggctaaa tttttggggt ctatgcttga
     3841 caaagcgagt taaaacctta gaatttaaga caggtacatt aagcccctgt ggtgaaatca
     3901 ttagcggtcg tcaaacctta atgctttcga ttgccgatat gtcagtatcg aagatctgta
     3961 ccccataaat tacggtaaag ccccaagcaa ttgcaagggg cttttatctt ttttaacaaa
     4021 aaaaatttat aaatcaggat tttataacac taaataatcc aaagtacatg agtaagtcat
     4081 gaccactcct gcgatgtgtg tagcactctg agtatccgta ttatatcagt gatgtcatat
     4141 acaaccacat aatgcggatg tatcactaac tccctagtat ctttctgtct gcctgtgcgc
     4201 cccatcagtc gattatcaac aagtaaatct gtcttttctt caattaaatc atcaatttca
     4261 atagctgcta tagggttgcg ttcttcaaga taatcataga tatttctacg atcttggctt
     4321 gcagccctta tgtcctcagg cgcgcacaaa acagcaggga agcagcgctt ttccgctgca
     4381 taaccctgct tcggggtcat tatagcgatt ttttcggtat atccatcctt tttcgcacga
     4441 tatacaggat tttgccaaag ggttcgtgta gactttcctt ggtgtatcca acggcgtcag
     4501 ccgggcagga taggtgaagt aggcccaccc gcgagcgggt gttccttctt cactgtccct
     4561 tattcgcacc tggcggtgct caacgggaat cctgctctgc gaggctggcc ggctaccgcc
     4621 ggcgtaacag atgagggcaa gcggatggct gatgaaacca agccaaccag gaagggcagc
     4681 ccacctatca aggtgtactg ccttccagac gaacgaagag cgattgagga aaaggcggcg
     4741 gcggccggca tgagcctgtc ggcctacctg ctggccgtcg gccagggcta caaaatcacg
     4801 ggcgtcgtgg actatgagca cgtccgcgag ctggcccgca tcaatggcga cctgggccgc
     4861 ctgggcggcc tgctgaaact ctggctcacc gacgacccgc gcacggcgcg gttcggtgat
     4921 gccacgatcc tcgccctgct ggcgaagatc gactctagct agaaatattt tatctgatta
     4981 ataagatgat cttcttgaga tcgttttggt ctgcgcgtaa tctcttgctc tgaaaacgaa
     5041 aaaaccgcct tgcagggcgg tttttcgaag gttctctgag ctaccaactc tttgaaccga
     5101 ggtaactggc ttggaggagc gcagtcacca aaacttgtcc tttcagttta gccttaaccg
     5161 gcgcatgact tcaagactaa ctcctctaaa tcaattacca gtggctgctg ccagtggtgc
     5221 ttttgcatgt ctttccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     5281 ggactgaacg gggggttcgt gcatacagtc cagcttggag cgaactgcct acccggaact
     5341 gagtgtcagg cgtggaatga gacaaacgcg gccataacag cggaatgaca ccggtaaacc
     5401 gaaaggcagg aacaggagag cgcacgaggg agccgccagg gggaaacgcc tggtatcttt
     5461 atagtcctgt cgggtttcgc caccactgat ttgagcgtca gatttcgtga tgcttgtcag
     5521 gggggcggag cctatggaaa aacggctttg ccgcggccct ctcacttccc tgttaaaccg
     5581 gaattctagg cccaaacgg
//
