LOCUS       sequence-8533-ex        6089 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6089)
  TITLE     Zebrafish Expression Plasmids
REFERENCE   2  (bases 1 to 6089)
  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: ZE49 pSPORT1-Sp5-l
FEATURES             Location/Qualifiers
     source          1..6089
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    650..671
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(686..691,692..709,710..716)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    724..740
                     /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     729..751
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     748..764
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     748..764
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     783..802
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        783..801
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1487..1516
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             1490..1519
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     promoter        complement(2898..2916)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2899..2916)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2924..2941)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2924..2940)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2933..2955)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3082..3537
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3169..3188)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3379..3400
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3564..3668
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3669..3737,3738..4529)
                     /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"
     primer_bind     complement(3887..3906)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4700..5288
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5189..5208
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5442..5459
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5566..5643
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(5644..6089,1..637)
                     /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     complement(5663..5682)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 tgtctctgac cagacaccca tcaacagtat tattttctcc catgaagacg gtacgcgact
       61 gggcgtggag catctggtcg cattgggtca ccagcaaatc gcgctgttag cgggcccatt
      121 aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat aaatatctca ctcgcaatca
      181 aattcagccg atagcggaac gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac
      241 catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca acgatcagat
      301 ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg cggatatctc
      361 ggtagtggga tacgacgata ccgaagacag ctcatgttat atcccgccgt taaccaccat
      421 caaacaggat tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc aactctctca
      481 gggccaggcg gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac
      541 cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
      601 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
      661 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
      721 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
      781 tctaatacga ctcactatag ggaaagctgg tacgcctgca ggtaccggtc cggaattccc
      841 gggtcgacgg acttattgcg acaggccctt cagtcgcact tcaaaaggaa gagacctggt
      901 ttgaaagacg ctgcttttga aaacgagacc cagaaaaaca ggagcaacag cgcggagtgg
      961 ccaacaactc tcacaggcga gaaaggacag gatttggaag agagaatcca aggattattg
     1021 atttttcagc gatatatctg aacgtacgca cctggctgtg taagggggtc aacaggatct
     1081 ttgcgcgcct cctcgttccc gcacactgag actttggcac ggctacgcac cgcgcgccgc
     1141 gcttcacgca ggtgtggata tcttgaaaca accgtgaaca catggctgca ttaaccttat
     1201 ccagagcgga caatttcctc cacaatttct tacaggaccg cactcccagc tcatctccag
     1261 aaagtggtcc aaacaccctc tcctttctgg ccaccacatg cagtcaggct tggcaggaag
     1321 gctcgcagct cccttacgag ggtccggtgg gctccgcatc cgcatgttcc agctctggag
     1381 caacgacgtg gccccgaact ccagcttaag cgctcaccag atgaccttca ccgtgcccaa
     1441 aatgcagttt cccagccaca tgcaacccac tctgggctcg cactcccatc accaccatca
     1501 ccaccatcat caccaccacg agcttcccct cactcctcca gcggagcctc cgtctgcata
     1561 ttcattcgag ctttccccgg ttaaaatgca agggaatgcg ccatactaca cgcagcacaa
     1621 cgccgtcagt caaaactttc caagcttcct ccaaaacccc tcaggaagag ctcacctgcc
     1681 tggaggtcac gttgaggatg gccagcagtg gtggagtctc ccccagggta acagcgctcc
     1741 ctccgggcac cctttctcct tgggacggca gctggttttg ggccatcagc ctcaaattgc
     1801 agcacttctg cagggaacct ccaaaggtct gttgagctct actcgtcgct gtcgccgatg
     1861 caaatgccca aactgccagt ccacaggaaa cggaggtgct gcattggagt ttggaaagaa
     1921 aagactacac atttgtcaca ttccagattg tgggaaggtt tataaaaaaa cctctcacct
     1981 caaagcgcac ctgcgctggc atgctggtga acggcccttc atttgcaact ggctcttctg
     2041 cgggaagagc ttcacgcgtt cagatgagct gcagaggcat ctccgtacac acaccgggga
     2101 gaagcgtttc ggctgtcagc agtgtggaaa acggtttatg cggagcgatc atctctccaa
     2161 gcatgtgaag acccatcaga gccggaagag ccggtccagt cagcctcctc acagcgggac
     2221 tgatgctctg ctgagcaaca tcaagagaga gtaaaagatg cacaagtact aaaaacaata
     2281 tgaacaagag ctaagagaga gaactttttt agagtctgga agtttggcag tggtgaaatg
     2341 gttgcgatgg gagcatgggc ctgtttctac tactgattgt ttcctgaaaa cactggagta
     2401 aagtgctgca ccaaaactga tgttataaag actgtaaaga gacttcaggg caaagcacaa
     2461 gtggtgtgcg agaggcattt tagtatgcct catgcccttg cacatttatt taaagacaat
     2521 ttaattcaaa atgagaaaaa agtattcgtt atttggtgaa actgtccttc ctttgtttac
     2581 attctgttgt ttgttttctc tattttgctt tagggaaaat tgatgcctta tgttgaaatc
     2641 ctgaattcag aacatttgcc aactaaattc ttgttttctt gcagagaatg gcactgctat
     2701 attcactcta aatccctata gtttcaaaac aagaaccaaa tgtatataat gtttatatag
     2761 cgcacagttc tataaagacc agcatgtgta ataataataa taaacacgat cgactgtgca
     2821 aatcaaaaaa aaaaaaaaag ggcggccgct ctagaggatc caagcttacg tacgcgtgca
     2881 tgcgacgtca tagctcttct atagtgtcac ctaaattcaa ttcactggcc gtcgttttac
     2941 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     3001 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     3061 gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     3121 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     3181 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     3241 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     3301 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     3361 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     3421 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     3481 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttcag
     3541 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     3601 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     3661 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     3721 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     3781 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     3841 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     3901 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     3961 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     4021 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     4081 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     4141 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     4201 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     4261 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     4321 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     4381 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     4441 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     4501 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     4561 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     4621 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     4681 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     4741 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     4801 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     4861 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     4921 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     4981 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     5041 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag cattgagaaa
     5101 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     5161 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     5221 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     5281 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     5341 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     5401 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     5461 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     5521 gcagagcttg caattcgcgc gcgaaggcga agcggcattt acgttgacac catcgaatgg
     5581 cgcaaaacct ttcgcggtat ggcatgatag cgcccggaag agagtcaatt cagggtggtg
     5641 aatgtgaaac cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc ttatcagacc
     5701 gtttcccgcg tggtgaacca ggccagccac gtttctgcga aaacgcggga aaaagtggaa
     5761 gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa
     5821 cagtcgttgc tgattggcgt tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt
     5881 gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta
     5941 gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc gcaacgggtc
     6001 agtgggctga tcattaacta tccgctggat gaccaggatg ccattgctgt ggaagctgcc
     6061 tgcactaatg ttccggcgtt atttcttga
//
