LOCUS       sequence-176792-        4503 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Increases expression of sperm whale myoglobin protein for making 
            apomyoglobin as a reagent for measuring rates of hemin dissociation 
            (as opposed to previous construct in pUC)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4503)
  TITLE     Sperm whale Mb 64Y68F in pVP80K
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4503)
  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            "Unpublished"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: Sperm whale Mb 64Y68F in pVP80K
FEATURES             Location/Qualifiers
     source          1..4503
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     84..102
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(126..140,141..146,147..163,164..170)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    146..162
                     /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)."
     protein_bind    178..194
                     /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     complement(810..828)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     protein_bind    complement(842..866)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      952..1038
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1131..2213)
                     /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="MKPVTLYVVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(2214..2291)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     complement(2336..2358)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2444..2584
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2499..2518
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2599..2616)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2770..3358)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2850..2869)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3535..4329)
                     /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="MLEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3646..3665)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4256..4275
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
       61 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt cttcacctcg
      121 agaaatcata aaaaatttat ttgctttgtg agcggataac aattataata gattcaattg
      181 tgagcggata acaatttcac actaaaaaac tcataaggta aggtatataa atggttctgt
      241 ctgaaggtga atggcagctg gttctgcatg tttgggctaa agttgaagct gacgtcgctg
      301 gtcatggtca ggacatcttg attcgactgt tcaaatctca tccggaaact ctggaaaaat
      361 tcgatcgttt caaacatctg aaaactgaag ctgaaatgaa agcttctgaa gatctgaaaa
      421 aatacggtgt taccttccta actgccctag gtgctatcct taagaaaaaa gggcatcatg
      481 aagctgagct caaaccgctt gcgcaatcgc atgctactaa acataagatc ccgatcaaat
      541 acctggaatt catctctgaa gcgatcatcc atgttctgca ttctagacat ccaggtaact
      601 tcggtgctga cgctcagggt gctatgaaca aagctctcga gctgttccgt aaagatatcg
      661 ctgctaagta caaagaactg ggttaccagg gttaatgatt aaacgaattc gagctcggta
      721 cccggggatc ctctagagtc gacctgcagg catgcaagct gatccggctg ctaacaaagc
      781 ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcaa actcgtttct
      841 cgttcagctt tcttgtacaa agtggtgatg gcgcgcctgt aggacgtcga cggtaccatc
      901 gatacgcgtt cgaagcttcg cctggggtaa tgactctcta gcttgaggca tcaaataaaa
      961 cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct
     1021 ctcctgagta ggacaaatcc gccctctaga ttacgtgcag tcgatgataa gctgtcaaac
     1081 atgagaattg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc tcactgcccg
     1141 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     1201 gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga gacgggcaac
     1261 agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc cacgctggtt
     1321 tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata acatgagctg
     1381 tcttcggtat cgtcgtatcc cactaccgag atatccgcac caacgcgcag cccggactcg
     1441 gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga
     1501 acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg
     1561 ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc
     1621 agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga
     1681 cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga gaaaataata
     1741 ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt agtgcaggca
     1801 gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag cccactgacg
     1861 cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct tcgttctacc
     1921 atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc cgcgacaatt
     1981 tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa cgactgtttg
     2041 cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat cgccgcttcc
     2101 actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc
     2161 tgataagaga caccggcata ctctgcgaca acgtataacg ttactggttt cacattcacc
     2221 accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt tttgcgccat
     2281 tcgatggtgt cggaacctag agctgcctcg cgcgtttcgg tgatgacggt gaaaacctct
     2341 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     2401 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt
     2461 cacgtagcga tagcggagtg tatactggct taactatgcg gcatcagagc agattgtact
     2521 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     2581 caggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     2641 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     2701 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     2761 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     2821 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     2881 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     2941 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     3001 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     3061 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     3121 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     3181 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     3241 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     3301 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     3361 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     3421 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     3481 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagcc taggtcagaa
     3541 gaactcgtca agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta
     3601 aagcacgagg aagcggtcag cccattcgcc gccaagctct tcagcaatat cacgggtagc
     3661 caacgctatg tcctgatagc ggtccgccac acccagccgg ccacagtcga tgaatccaga
     3721 aaagcggcca ttttccacca tgatattcgg caagcaggca tcgccatgag tcacgacgag
     3781 atcctcacca tccggcatac gcgccttgag cctggcgaac agttcggctg gcgcgagccc
     3841 ctgatgctct tcgtccagat catcctgatc gacaagaccg gcttccatcc gagtgcgtgc
     3901 tcgctcgatg cgatgtttcg cttggtggtc gaatgggcag gtagccggat caagcgtatg
     3961 cagccgccgc attgcatcag ccatgatgga tactttctcg gcaggagcaa ggtgagatga
     4021 caggagatcc tgccccggca cttcgcccaa tagcagccag tcccttcccg cttcagtgac
     4081 aacgtcgagc acagctgcgc aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc
     4141 ctcgtcctgc agttcattca gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg
     4201 cccctgcgct gacagccgga acacggcggc atcagagcag ccgattgtct gttgtgccca
     4261 gtcatagccg aatagcctct ccacccaagc ggccggagaa cctgcgtgca atccatcttg
     4321 ttcaagcatg cgaaacgacc gtcatcctgt ctcttgatca gatcttgatc ccctgcgcca
     4381 tcagatcctt ggcggcaaga aagccatcca gtttactttg cagggcttcc caaccttacc
     4441 agagggcgcc ccagctggca attcttttga agctcacgct gccgcaagca ctcagggcgt
     4501 acg
//
