LOCUS       Exported                8507 bp ds-DNA     circular SYN 08-MAR-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8507)
  AUTHORS   Bioproduction engineering laboratory
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Mar 10, 2022 from SnapGene 5.2.4
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8507
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        96..200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             201..1061
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1232..1820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        2317..3254
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
     CDS             3261..3515
                     /codon_start=1
                     /gene="MF-alpha-1"
                     /product="N-terminal secretion signal from S. cerevisiae 
                     alpha-factor"
                     /label=alpha-factor secretion signal
                     /note="Cleavage by the Kex2 protease occurs after the 
                     dibasic KR sequence."
                     /translation="MRFPSIFTAVLFAASSALAAPVNTTTEDETAQIPAEAVIGYLDLE
                     GDFDVAVLPFSNSTNNGLLFINTTIASIAAKEEGVSLDKR"
     CDS             3531..6056
                     /codon_start=1
                     /label=AaBGL1
                     /translation="DELAFSPPFYPSPWANGQGEWAEAYQRAVAIVSQMTLDEKVNLTT
                     GTGWELEKCVGQTGGVPRLNIGGMCLQDSPLGIRDSDYNSAFPAGVNVAATWDKNLAYL
                     RGQAMGQEFSDKGIDVQLGPAAGPLGRSPDGGRNWEGFSPDPALTGVLFAETIKGIQDA
                     GVVATAKHYILNEQEHFRQVAEAAGYGFNISDTISSNVDDKTIHEMYLWPFADAVRAGV
                     GAIMCSYNQINNSYGCQNSYTLNKLLKAELGFQGFVMSDWGAHHSGVGSALAGLDMSMP
                     GDITFDSATSFWGTNLTIAVLNGTVPQWRVDDMAVRIMAAYYKVGRDRLYQPPNFSSWT
                     RDEYGFKYFYPQEGPYEKVNHFVNVQRNHSEVIRKLGADSTVLLKNNNALPLTGKERKV
                     AILGEDAGSNSYGANGCSDRGCDNGTLAMAWGSGTAEFPYLVTPEQAIQAEVLKHKGSV
                     YAITDNWALSQVETLAKQASVSLVFVNSDAGEGYISVDGNEGDRNNLTLWKNGDNLIKA
                     AANNCNNTIVVIHSVGPVLVDEWYDHPNVTAILWAGLPGQESGNSLADVLYGRVNPGAK
                     SPFTWGKTREAYGDYLVRELNNGNGAPQDDFSEGVFIDYRGFDKRNETPIYEFGHGLSY
                     TTFNYSGLHIQVLNASSNAQVATETGAAPTFGQVGNASDYVYPEGLTRISKFIYPWLNS
                     TDLKASSGDPYYGVDTAEHVPEGATDGSPQPVLPAGGGSGGNPRLYDELIRVSVTVKNT
                     GRVAGDAVPQLYVSLGGPNEPKVVLRKFDRLTLKPSEETVWTTTLTRRDLSNWDVAAQD
                     WVITSYPKKVHVGSSSRQLPLHAALPKVQ"
     terminator      6063..6396
                     /label=AOX1t
     misc_feature    6409..7008
                     /label=T38473
     CDS             complement(7026..7820)
                     /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"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agcattgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2101 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     2161 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     2221 accatgatta cgccaagctt gcggccgcgg atcccatgtt ggtattgtga aatagacgca
     2281 gatcgggaac actgaaaaat aacagttatt attcgagatc taacatccaa agacgaaagg
     2341 ttgaatgaaa cctttttgcc atccgacatc cacaggtcca ttctcacaca taagtgccaa
     2401 acgcaacagg aggggataca ctagcagcag accgttgcaa acgcaggacc tccactcctc
     2461 ttctcctcaa cacccacttt tgccatcgaa aaaccagccc agttattggg cttgattgga
     2521 gctcgctcat tccaattcct tctattaggc tactaacacc atgactttat tagcctgtct
     2581 atcctggccc ccctggcgag gttcatgttt gtttatttcc gaatgcaaca agctccgcat
     2641 tacacccgaa catcactcca gatgagggct ttctgagtgt ggggtcaaat agtttcatgt
     2701 tccccaaatg gcccaaaact gacagtttaa acgctgtctt ggaacctaat atgacaaaag
     2761 cgtgatctca tccaagatga actaagtttg gttcgttgaa atgctaacgg ccagttggtc
     2821 aaaaagaaac ttccaaaagt cggcataccg tttgtcttgt ttggtattga ttgacgaatg
     2881 ctcaaaaata atctcattaa tgcttagcgc agtctctcta tcgcttctga accccggtgc
     2941 acctgtgccg aaacgcaaat ggggaaacac ccgctttttg gatgattatg cattgtctcc
     3001 acattgtatg cttccaagat tctggtggga atactgctga tagcctaacg ttcatgatca
     3061 aaatttaact gttctaaccc ctacttgaca gcaatatata aacagaagga agctgccctg
     3121 tcttaaacct tttttttatc atcattatta gcttactttc ataattgcga ctggttccaa
     3181 ttgacaagct tttgatttta acgactttta acgacaactt gagaagatca aaaaacaact
     3241 aattattcga aacgactagt atgagatttc cttcaatttt tactgcagtt ttattcgcag
     3301 catcctccgc attagctgct ccagtcaaca ctacaacaga agatgaaacg gcacaaattc
     3361 cggctgaagc tgtcatcggt tacttagatt tagaagggga tttcgatgtt gctgttttgc
     3421 cattttccaa cagcacaaat aacgggttat tgtttataaa tactactatt gccagcattg
     3481 ctgctaaaga agaaggggta tctttggata aaagagacgt cgacgaattc gatgaactgg
     3541 cgttctctcc tcctttctac ccctctccgt gggccaatgg ccagggagag tgggcggaag
     3601 cctaccagcg tgcagtggcc attgtatccc agatgactct ggatgagaag gtcaacctga
     3661 ccaccggaac tggatgggag ctggagaagt gcgtcggtca gactggtggt gtcccaagac
     3721 tgaacatcgg tggcatgtgt cttcaggaca gtcccttggg aattcgtgat agtgactaca
     3781 attcggcttt ccctgctggt gtcaacgttg ctgcgacatg ggacaagaac cttgcttatc
     3841 tacgtggtca ggctatgggt caagagttca gtgacaaagg aattgatgtt caattgggac
     3901 cggccgcggg tcccctcggc aggagccctg atggaggtcg caactgggaa ggtttctctc
     3961 cagacccggc tcttactggt gtgctctttg cggagacgat taagggtatt caagacgctg
     4021 gtgtcgtggc gacagccaag cattacattc tcaatgagca agagcatttc cgccaggtcg
     4081 cagaggctgc gggctacgga ttcaatatct ccgacacgat cagctctaac gttgatgaca
     4141 agaccattca tgaaatgtac ctctggccct tcgcggatgc cgttcgcgcc ggcgttggcg
     4201 ccatcatgtg ttcctacaac cagatcaaca acagctacgg ttgccagaac agttacactc
     4261 tgaacaagct tctgaaggcc gagctcggct tccagggctt tgtgatgtct gactggggtg
     4321 ctcaccacag tggtgttggc tctgctttgg ccggcttgga tatgtcaatg cctggcgata
     4381 tcaccttcga ttctgccact agtttctggg gtaccaacct gaccattgct gtgctcaacg
     4441 gtaccgtccc gcagtggcgc gttgacgaca tggctgtccg tatcatggct gcctactaca
     4501 aggttggccg cgaccgcctg taccagccgc ctaacttcag ctcctggact cgcgatgaat
     4561 acggcttcaa gtatttctac ccccaggaag ggccctatga gaaggtcaat cactttgtca
     4621 atgtgcagcg caaccacagc gaggttattc gcaagttggg agcagacagt actgttctac
     4681 tgaagaacaa caatgccctg ccgctgaccg gaaaggagcg caaagttgcg atcctgggtg
     4741 aagatgctgg atccaactcg tacggtgcca atggctgctc tgaccgtggc tgtgacaacg
     4801 gtactcttgc tatggcttgg ggtagcggca ctgccgaatt cccatatctc gtgacccctg
     4861 agcaggctat tcaagccgag gtgctcaagc ataagggcag cgtctacgcc atcacggaca
     4921 actgggcgct gagccaggtg gagaccctcg ctaaacaagc cagtgtctct cttgtatttg
     4981 tcaactcgga cgcgggagag ggctatatct ccgtggacgg aaacgagggc gaccgcaaca
     5041 acctcaccct ctggaagaac ggcgacaacc tcatcaaggc tgctgcaaac aactgcaaca
     5101 acaccatcgt tgtcatccac tccgttggac ctgttttggt tgacgagtgg tatgaccacc
     5161 ccaacgttac tgccatcctc tgggcgggct tgcctggcca ggagtctggc aactccttgg
     5221 ctgacgtgct ctacggccgc gtcaacccgg gcgccaaatc tccattcacc tggggcaaga
     5281 cgagggaggc gtacggggat taccttgtcc gtgagctcaa caacggcaac ggagctcccc
     5341 aagatgattt ctcggaaggt gttttcattg actaccgcgg attcgacaag cgcaatgaga
     5401 ccccgatcta cgagttcgga catggtctga gctacaccac tttcaactac tctggccttc
     5461 acatccaggt tctcaacgct tcctccaacg ctcaagtagc cactgagact ggcgccgctc
     5521 ccaccttcgg acaagtcggc aatgcctctg actacgtgta ccctgaggga ttgaccagaa
     5581 tcagcaagtt catctatccc tggcttaatt ccacagacct gaaggcctca tctggcgacc
     5641 cgtactatgg agtcgacacc gcggagcacg tgcccgaggg tgctactgat ggctctccgc
     5701 agcccgttct gcctgccggt ggtggctctg gtggtaaccc gcgcctctac gatgagttga
     5761 tccgtgtttc ggtgacagtc aagaacactg gtcgtgttgc cggtgatgct gtgcctcaat
     5821 tgtatgtttc ccttggtgga cccaatgagc ccaaggttgt gttgcgcaaa ttcgaccgcc
     5881 tcaccctcaa gccctccgag gagacggtgt ggacgactac cctgacccgc cgcgatctgt
     5941 ctaactggga cgttgcggct caggactggg tcatcacttc ttacccgaag aaggtccatg
     6001 ttggtagctc ttcgcgtcag ctgccccttc acgcggcgct cccgaaggtg caatgactcg
     6061 agccttagac atgactgttc ctcagttcaa gttgggcact tacgagaaga ccggtcttgc
     6121 tagattctaa tcaagaggat gtcagaatgc catttgcctg agagatgcag gcttcatttt
     6181 tgatactttt ttatttgtaa cctatatagt ataggatttt ttttgtcatt ttgtttcttc
     6241 tcgtacgagc ttgctcctga tcagcctatc tcgcagctga tgaatatctt gtggtagggg
     6301 tttgggaaaa tcattcgagt ttgatgtttt tcttggtatt tcccactcct cttcagagta
     6361 cagaagatta agtgagaagt tcgtttgtgc aagcttagat cttctagaga caataagaag
     6421 aaaaaaaaag aaaagcggtg ggggagggat tattaaataa ggattatgta accccagggt
     6481 accgttctat acatatttaa ggattattta ggacaatcga tgaaatcggc atcaaactgg
     6541 atgggagtat agtgtccgga taatcggata aatcatcttg cgaggagccg cttggttggt
     6601 tggtgagagg agtgaaatat gtgtctcctc acccaagaat cgcgatatca gcaccctgtg
     6661 ggggacacta ttggcctccc tcccaaacct tcgatgtggt agtgctttat tatattgatt
     6721 acattgatta catagctaaa ccctgcctgg ttgcaagttg agctccgaat tccaatatta
     6781 gtaaaatgcc tgcaagataa cctcggtatg gcgtccgacc ccgcttaatt attttaactc
     6841 ctttccaacg aggacttcgt aatttttgat tagggagttg agaaacgggg ggtcttgata
     6901 cctcctcgat ttcagatccc accccctctc agtcccaagt gggacccccc tcggccgtga
     6961 aatgcgcgca ctttagtttt tttcgcatgt aaacgccggt gtccgtcatc tagagaattc
     7021 cccgctcaga agaactcgtc aagaaggcga tagaaggcga tgcgctgcga atcgggagcg
     7081 gcgataccgt aaagcacgag gaagcggtca gcccattcgc cgccaagctc ttcagcaata
     7141 tcacgggtag ccaacgctat gtcctgatag cggtccgcca cacccagccg gccacagtcg
     7201 atgaatccag aaaagcggcc attttccacc atgatattcg gcaagcaggc atcgccatgg
     7261 gtcacgacga gatcctcgcc gtcgggcatg cgcgccttga gcctggcgaa cagttcggct
     7321 ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat cgacaagacc ggcttccatc
     7381 cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca ggtagccgga
     7441 tcaagcgtat gcagccgccg cattgcatca gccatgatgg atactttctc ggcaggagca
     7501 aggtgagatg acaggagatc ctgccccggc acttcgccca atagcagcca gtcccttccc
     7561 gcttcagtga caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc cagccacgat
     7621 agccgcgctg cctcgtcctg cagttcattc agggcaccgg acaggtcggt cttgacaaaa
     7681 agaaccgggc gcccctgcgc tgacagccgg aacacggcgg catcagagca gccgattgtc
     7741 tgttgtgccc agtcatagcc gaatagcctc tccacccaag cggccggaga acctgcgtgc
     7801 aatccatctt gttcaatcat atggtttcta tattatcttt gtactaaaga gcaattgata
     7861 atgtgcgaga aaaactggtc cttatatgcc gtttgcagca ctccctcccg aactttacga
     7921 aaagtcgtgc gccacctgat tttcatcacg ccaaaaacct acacgtatga ctactccggg
     7981 ccagtgttca ccacgagcta tatagtgtta attaattacc ttattggtta gctctgcatg
     8041 taagggtggt gtgagccggg aattgggtct actctagcgt tcagtaaggt gatataaagc
     8101 tctgtagaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     8161 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     8221 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg
     8281 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac
     8341 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
     8401 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     8461 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcga
//
